bluuebunny/crossref_metadata_2025 huggingface dataset:
Lymphocyte stimulation: a rapid multiparameter analysis.
Authors: Darzynkiewicz Z , Traganos F , Sharpless T , Melamed M R
Published: May 2006

Several parameters of stimulation of individual lymphocytes are measured simultaneously by flow-cytofluorometry after differential staining of cellular DNA and RNA with the metachromatic fluorescent dye acridine orange. The method provides a means of analyzing the progression of stimulated cells through the cell cycle (G1, S and G2 + M), in addition to measuring the accumulation of RNA per cell; the RNA parameter is useful in distinguishing G1 from G0 cells. The multiparameter analysis may be of unique value in discriminating cases where the transcriptional and proliferative responses of lymphocytes to stimulants are not correlated.
Organization of DNA in chromatin.
Authors: Sobell H M , Tsai C C , Gilbert S G , Jain S C , Sakore T D
Published: May 2006

Conformational changes in DNA that accompany drug intercalation have led us to ask if DNA first bends or "kinks" to accept an intercalative drug or dye. Kinking is made possible by altering the normal C2' endo deoxyribose sugar ring puckering in B DNA to a mixed sugar puckering pattern of the type C3' and partially unstacking base-pairs. A kinking scheme such as this would require minimal stereochemical rearrangement and would also involve small energies. This has prompted us to ask more generally if a conformational change such as this could be used by proteins in their interactions with nucleic acids. In this papter we describe an interesting superhelical DNA structure formed by kinking DNA every 10 base-pairs. The structure may be used in the organization of DNA in chromatin.
Autoregulation of simian virus 40 gene A by T antigen.
Authors: Reed S I , Stark G R , Alwine J C
Published: May 2006

During lytic infection by simian virus 40, gene A is transcribed into early RNA, which is translated into A protein (T antigen). Both the rate of synthesis and the intracellular amount of early RNA are higher in cells infected by temperature-sensitive A (tsA) mutants than in cells infected by wild-type virus. These differences are observed at permissive temperature (32 degrees) and are amplified greatly after a shift to restrictive temperature (41 degrees). For example, at 32 degrees cells infected by tsA mutants synthesize early RNA approximately twice as fast as cells infected by wild-type virus. After the shift to 41 degrees, the rate of synthesis in the tsA infection increases to 15 times the rate in the wild-type infection. In contrast, cells infected by tsA mutants do not overproduce late RNA. We suggest that the A protein regulates its own synthesis by negative feedback control of gene A transcription.
Isolation of a set of hybrid lac repressors made in vitro between normal lac repressor and its homogeneous tryptic core.
Authors: Geisler N , Weber K
Published: May 2006

Lactose repressor can be renatured from 8 M guanidine-HCl solution. The renatured repressor is tetrameric and shows DNA binding activity. Thus it becomes possible to obtain hybrid tetramers in vitro between normal repressor and repressor defective in DNA binding by simultaneous denaturation and renaturation. In order to facilitate the separation of the different hybrids, we have used a lac repressor derivative that does not bind DNA, which is missing the amino-terminal 59 residues of the polypeptide chain (homogeneous tryptic core). The hybrids resulting from the mixed renaturation of homogeneous tryptic core and normal repressor can be separated by electrophoresis on Cellogel. The hybrids have been recovered, and a preliminary characterization of their DNA-binding properties is reported.
The identification of the mot gene product with Escherichia coli-lambda hybrids.
Authors: Silverman M , Matsumura P , Simon M
Published: May 2006

Molecular cloning techniques were used to construct lambda-E. coli hybrid bacteriophage carrying genes involved in bacterial flagellar motility (mot) and chemotaxis (cheA). A series of hybrid bacteriophage without each of these genes was also prepared.When paralyzed mutants of E. coli were infected with lambda that carried the mot gene, the ability of the bacterium to swim was rapidly restored. The restoration of motility was the result of the synthesis and insertion into the cell membrane of a protein with an apparent molecular weight of 31,000 (the Mot protein). Another polypeptide with a mobility on acrylamide gel electrophoresis which corresponded to a molecular weight of 39,000 was associated with the cheA gene. The presence of this polypeptide alone was not sufficient to restore chemotactic activity to mutant cheA strains. It was suggested that only a portion of the cheA gene was cloned, and thus the 39,000 protein may be a partial product of the cheA gene, or the product of a second mot gene.
Differences among sulfated proteoglycans synthesized in nonchondrogenic cells, presumptive chondroblasts, and chondroblasts.
Authors: Okayama M , Pacifici M , Holtzer H
Published: May 2006

The sulfated proteoglycans synthesized by definitive chondroblasts in cultured 10-day chick vertebral or epiphyseal cartilages were characterized by their sedimentation profile in a sucrose gradient and their susceptibility to chondroitinase ABC (EC 4.2.2.4; chondroitin ABC lyase). These sulfated proteoglycans were indistinguishable from those synthesized by definitive chondroblasts that emerge from older cultures of somites plus notochord or in older cultures of limb buds. The sulfated proteoglycans of these definitive chondroblasts are readily distinguished from those synthesized by their mother cells, the presumptive chondroblasts, or those synthesized by dedifferentiated or bromodeoxyuridine-suppressed chondroblasts. However, the sulfated proteoglycans synthesized by presumptive chondroblasts or by dedifferentiated or bromodeoxyuridine-suppressed chondroblasts cannot be dintinguished by these techniques from those synthesized by (i) blastodisc cells, (ii) fibroblasts, (iii) spinal cord cells, or (iv) skeletal, cardiac, or smooth muscle cells. Addition of glycosaminoglycans or collagen to the medium did not induce somite or limb presumptive chondroblasts to synthesize the chondroblast-unique sulfated proteoglycans. Cells moving from the presumptive chondroblast compartment into the chondroblast compartment acquire not only the option to initiate the synthesis of chondroblast-unique collagen chains, but also the capacity to synthesize chondroblast-unique sulfated proteoglycans.
Selective cannibalism in the rotifer asplanchna sieboldi: contact recognition of morphotype and clone.
Authors: Gilbert J J
Published: May 2006

Populations of A. sieboldi reproduce primarily by diploid femal parthenogenesis. Females may exhibit a pronounced, nongenetic polymorphism which is controlled by the diet. The cruciform and especially the campanulate morphotypes are larger than the saccate morphotype and are cannibalistic. Direct observations of various predator-prey interactions between the different morphotypes of two taxonomically distinct clones show that the feeding responses of cannibals after actual prey contact are predictable and extremely selective. Companulates from clone B respond regularly to saccate but only rarely to cruciform or campanulate clonemates; the same predators typically respond to cruciforms and campanulates from clone C. Cruciforms from clone C rarely respond to saccate and cruciform clonemates but readily attack all morphotypes from clone B. In contrast, campanulates from clone C show no selectivity. Morphotype- and clone-specific recognition responses, which are mediated by coronal contact chemoreceptors, permit efficient prey discrimination and protection withotu involving prey handling or development of energy-demanding, defensive structures, such as the body-wall outgrowths of the cruciform morphotype. Selective feeding of cannibals may increase the fitness of a clone, both by effecting a more adaptive distribution of its morphotypes and by enhancing its ability to compete directly and indirectly with cooccurring clones. The magnitude of cannibalism in this rotifer may be dependent upon a complex suite of heterogeneous predator-prey interactions and greatly affected by shifting densities and distributions of different clones and female morphotypes.
Loss of mouse chromosomes in somatic cell hybrids between HT-1080 human fibrosarcoma cells and mouse peritioneal macrophages.
Authors: Croce C M
Published: May 2006

Somatic cell hybrids between mouse peritioneal macrophages and HT-1080 human fibrosarcoma cells lose mouse chromosomes and retain the entire complement of human chromosomes. In contrast, somatic cell hybrids between cells derived from two different mouse continuous cell lines and HT-1080 human cells were found to lose human chromosomes preferentially. Loss of mouse chromosomes is not a general property of hybrids between mouse macrophages and transformed human cells; the hybridization of mouse macrophages with cells derived from five different human fibroblast lines transformed by simian virus 40 resulted in the production of hybrid clones that preferentially lost human chromosomes.
Marked hydrophobicity of the NH2-terminal extra piece of immunoglobulin light-chain precursors: possible physiological functions of the extra piece.
Authors: Schechter I , Burstein Y
Published: May 2006

mRNAs coding for mouse immunoglobulin light chains direct the cell-free synthesis of precursors in which extra peptide segments precede the NH2-termini of the mature proteins. The abundance (18-30%) of leucine residues in the extra piece indicates that it is quite hydrophobic [Schechter and Burstein (1976) Biochem. Biophys, Res. Commun. 68, 489]. Accordingly, we have determined the positions of all hydrophobic residues by sequencing two k-type light (L)-chain precursors that were labeled with: [3H]Ala, [3H]Val, [3H]Leu, [3H]Ile, [3H]Thr, [3H]Pro, [3H]Phe, [3H]Tyr, [3H]Trp, [35S]Met, and [35S]Cys. The partial sequences (and sizes) of the extra pieces obtained are: in MOPC-321 precursor, Met-X-Thr-X-Thr-Leu-Leu-Leu-Trp-Val-Leu-Leu-Leu-Trp-Val-Pro-X-X-Thr-X-(20 residues; X is unknown); in MOPC-41 precursor, Met-X-Met-X-Ala-Pro-Ala-X-Ile-Phe-X-Phe-Leu-Leu-Leu-Leu-Phe-Pro-X-Thr-X-Cys- (22 residues). Despite the fact that these extra pieces differ extensively in sequence (68%), both of them are highly enriched with hydrophobic residues (75% in MOPC-321, 73% in MOPC-41). This marked hydrophobicity suggests that the extra piece favors interaction of the precursor with cell membranes, in a manner similar to the function of the "hydrophobic domain" of membrane-bound proteins (e.g., glycophorin). We propse that the hydrophobic extra piece directs most precursor molecules to the endoplasmic reticulum, where they are cleaved to yield mature L chain destined for scretion; a few precursor molecules escape cleavage and are embedded in the cell surface to serve as the antigen-recognizing receptor. The probability that the Leu-Leu-Leu-Trp-Val sequence occurs by change is 1.6 X 10(-8). Therefore, the data provide evidnece for duplication of a short DNA segment in the structural gene coding for the MOPC-321 precurosr. Duplication with inversion is also indicated from inverted repetition of the Phe-Lue-Leu sequence in the extra piece of the MPOC-41 precursor.
Cell surface antigens of human malignant melanoma: mixed hemadsorption assays for humoral immunity to cultured autologous melanoma cells.
Authors: Carey T E , Takahashi T , Resnick L A , Oettgen H F , Old L J
Published: May 2006

We studied how frequently patients with malignant melanoma have specific antibody to cell surface antigens of cultured autologous melanoma cells as demonstrated by mixed hemadsorption assays. Of 35 patients studied over periods ranging from 1 to 36 months with Stage II, III, and IV disease, two showed consistent and high titered reactivity against autologous melanoma cells, two showed less consistent and intermediate reactivity, seven showed sporatic, low titered reactivity, and the remainder were consistently negative. A detailed analysis was carried out with the sera of one patient with sufficiently high titer against autologous melanoma cells. By direct tests and by absorption analysis with a variety of melanoma and nonmelanoma cell lines which included autologous fibroblasts, the antigen could not be demonstrated on any cell type other than the autologous melanoma.
Effects of pressure on visible spectra of complexes of myoglobin, hemoglobin, cytochrome c, and horse radish peroxidase.
Authors: Ogunmola G B , Zipp A , Chen F , Kauzmann W
Published: May 2006

The spectra of the ferric form of most heme proteins [metmyoglobin, methemoglobin, horse radish peroxidase (EC 1.11.1.7), and ferricytochrome c at pH 1.5] are converted from high-spin (open crevice) structure to low-spin (closed crevice) form under pressure. Pressures up to 8000 kg/cm2 (780 MPa) have no effect on the spectra of high-spin ferro- and ferricytochrome c, which have a closed crevice structure at pH 7.0. Spectra of deoxy-ferromyoglobin and deoxy-ferrohemoglobin are reduced in intensity, but pressure does not change the positions of the absorption maxima. Cyanide ion prevents pressure-induced spectral changes in metmyoglobin and methemoglobin up to 8000 kg/cm2. Carbon monoxide (with a high affinity for the ferro heme iron) has a similar effect on ferromyoglobin and ferrohemoglobin. The pressure required to cause spectral changes in the heme proteins falls in the order, cytochrome c (pH 7.0) greater than horse radish peroxidase greater than myoglobin greater than hemoglobin. We have calculated a volume change of --50 cm3/mol associated with the configurational change accompanying the reformation of the iron-methionine bond in cytochrome c at low pH.
A novel prostaglandin is the major product of arachidonic acid metabolism in rabbit heart.
Authors: Isakson P C , Raz A , Denny S E , Pure E , Needleman P
Published: May 2006

The prostaglandins (PGs) released from the heart have generally been characterized as resembling PGE by bioassay techniques. The major PG formed from [14C]arachidonate (C20:4) by the isolated perfused rabbit heart has chromatographic mobility similar to that of PGE2 in most solvent systems. However, additional analysis of this radioactive "PGE" peak suggests that two substances are formed by the heart and migrate like PGE2: one has chemical properties similar to those of authentic PGE2 and the other is a novel PG. The unknown compound is the major PG formed by the heart from either exogenous arachidonate or hormonal stimulation of PG biosynthesis. The novel PG produced by the heart may be identical with either 6(9)-oxy-PGF or 6-keto-PGF1 alpha.
Actin microheterogeneity in chick embryo fibroblasts.
Authors: Rubenstein P A , Spudich J A
Published: May 2006

Secondary chick embryo fibroblasts contain three distinct actin species--alpha, beta, and lambda, in the approximate ratio of 1:6:3--with the same molecular weights but different isoelectric points. The most acidic of these components, alpha, comigrates on isoelectric focusing gels with the major actin of cardiac and skeletal muscle, while lambda, the most basic of the actins, comigrates with smooth muscle actin. The three components have overlapping methionine-containing tryptic peptides. All three actins are found to be present in actomyosin and cytoskeleton preparations from chick embryo fibroblasts. Identification of alpha-actin as the major actin from sarcomere-containing cells is confirmed by comparing embryonic chicken pre- and post-fusion myoblast cultures. Following myoblast fusion, the relative amount of alpha-actin increases until it changes from a minor actin component to the predominant actin species in the culture.
Intramolecular general base-catalyzed ester hydrolyses by the imidazolyl group.
Authors: Komiyama M , Roesel T R , Bender M L
Published: May 2006

Intramolecular general base catalysis by the imidazolyl group was found in the hydrolyses of endo-5-[4;(5')-imidazolyl]-bicyclo[2.2.1]hept-endo-2-yl trans-cinnamate and endo-5-[4'(5')-imidazolyl]bicyclo[2.2.2]oct-endo-2-yl trans-cinnamate in which the imidazolyl and trans-cinnamoyl groups are bound in close proximity to each other by rigid bicyclic rings. The rate constants for the intramolecular general base-catalyzed hydrolyses at 60 degrees are 6.4 X 10(-7) sec-1 for the former and 1.8 X 10(-7) sec-1 for the latter and the deuterium oxide solvent isotope effects are 3.0 for both. On the other hand, no intramolecular catalytic participation of the imidazolyl group was observed in the hydrolyses of the endo-exo isomers, exo- 5-[4'(5')-imidazolyl]bicyclo[2.2.1]hept-endo-2-yl trans-cinnamate and endo-5[4'(5')-imidazolyl]bicyclo[2.2.2]oct-exo-2-yl trans-cinnamate, in which the imidazolyl groups are located far from the trans-cinnamoyl groups. Intramolecular general base-catalyzed hydrolyses by the imidazolyl groups in endo-5[4'(5')-imidazolyl]bicyclo[2.2.1]hept-endo-2-yl trans-cinnamate and endo-5-[4'(5')-imidazolyl]bicyclo[2.2.2]oct-endo-2-yl trans-cinnamate can serve as models of serine esterase-catalyzed hydrolyses.
A mutant of Escherichia coli showing constitutive expression of the lysogenic induction and error-prone DNA repair pathways.
Authors: Mount D W
Published: May 2006

A mutant of E. coli (designated the STS mutant) has been isolated in which the phage induction and error-prone DNA repair pathways appear to be expressed constitutively without the cells having received an inducing signal. Phage lambda was not able to lysogenize this mutant, whereas a noninducible mutant of lambda, lambdacIind-, known to synthesize a repressor that is insensitive to the induction mechanism, lysogenized it normally. This result suggested that normal phage repressor was synthesized in the STS mutant but was then inactivated by the induction mechanism. The STS strain also had mutator characteristics, and showed spontaneous, error-prone repair of UV-damaged phage lambda. Derived from a lexA tif sfiA parent strain, the STS mutant carried an additional mutation spr at the lexA locus that resulted in a high level of expression of the induction pathways. The properties of this and related strains provide additional evidence that induction of phage and induction of error-prone DNA repair occur by a similar mechanism, and further suggest a model for the regulation of these pathways.
Immunological defect and its correction in the osteopetrotic mutant rat.
Authors: Milhaud G , Labat M L , Parant M , Damais C , Chedid L
Published: May 2006

Congenital osteopetrosis in the mutant rat "op" is accompanied by early atrophy of the thymus gland. The response of thymocytes to concanavalin A and phytohemagglutinin P was greatly diminished at age 28 days, even before pronounced thymic atrophy could be detected. The response of spleen cells to mitogens that stimulate thymus-derived (T) and bone-marrow-derived (B) cells was diminished as early as 35 days of age. A single injection of a suspension of normal bone marrow, which cures osteopetrosis, increased thymus weight and restored the normal responses of both thymus and spleen to various mitogens. These results add further support to the hypothesis that the thymus is involved in the pathogenesis of osteopetrosis.
Carcass components at first estrus of rats on high-fat and low-fat diets: body water, protein, and fat.
Authors: Frisch R E , Hegsted D M , Yoshinaga K
Published: May 2006

Carcass analysis data show that weanling rats fed high-fat (HF) and low-fat (LF) diets until the day of first estrus had similar body compositions at estrus, although the HF rats had estrus significantly earlier and at a lighter body weight (P less than 0.01) than the LF-diet rats. Total water as percent of wet weight, protein as percent of wet weight, and the water/protein ratios of the HF- and LF-diet rats did not differ significantly, whereas the absolute amounts of body water and protein of the two diet groups differed significantly (P less than 0.01), in accord with means (+/- SEM) of total body water as percent of wet weight were 66.2 +/- 0.3% for the HF rats and 66.4 +/- 0.3% for the LF-diet rats, whereas the mean absolute total body water for the two diet groups was 69.7 +/- 2.2 g and 81.1 +/- 2.4 g, respectively (P less than 0.01). Carcass fat of the HF rats, 15.6 +/- 1.0 g, was identical with that of the LF-diet rats, 15.6 +/- 0.9 g. The HF rats were therefore relatively fatter, 14.6 +/- 0.6 wet weight %, than the LF-diet rats, 12.6 +/- 0.4 wet weight % (0.05 greater than P greater than 0.01). The percentage of water in the fat-free wet weight of the HF rats, 77.5 +/- 0.3%, was significantly (P less than 0.01) greater than that of the LF-diet rats, 76.1 +/- 0.3%. These data and the high percentages of total water of body weight show that at first estrus the HF- and LF-diet rats had not as yet attained an adult body composition, similar to the human female at menarche. Within each diet group, the percentages of total water/body weight, protein/wet weight and fat/wet weight, or fat/dry weight, did not change significantly with increasing age of estrus, whereas each absolute carcass component--body water, protein, and fat--increased significantly with increasing age of estrus. The carcass data support G.C. Kennedy's hypothesis that a metabolic signal, related to fat stores, is a signal for puberty in the rat and are in accord with the hypothesis that a critical body composition of fatness is necessary for estrus in the rat, as in the human female. The greater relative fatness of the HF-diet rats may be associated with higher levels of estrogen in the HF rats than in the LF-diet rats.
Selective and accurate transcription of the Xenopus laevis 5S RNA genes in isolated chromatin by purified RNA polymerase III.
Authors: Parker C S , Roeder R G
Published: May 2006

Chromatin isolated from immature oocytes was found to contain an endogenous RNA polymerase activity (RNA nucleotidyltransferase; nucleoside triphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) that synthesizes predominately 5S RNA. However, the levels of total RNA synthesis and 5S RNA synthesis in chromatin were each stimulated 10- to 50-fold by an exogenous RNA polymerase III purified from X. laevis oocytes. The 5S genes in chromatin were transcribed by the exogenous enzyme in a highly selective (3000-fold above random) and predominately asymmetric fashion. A significant fraction of 5S RNA sequences were also found in a discrete transcript, approximately 5S in size. Total RNA synthesis was significantly stimulated when chromatin was transcribed by oocyte RNA polymerase I, murine RNA polymerase II, and low levels of Escherichia coli RNA polymerase. However, these enzymes did not significantly stimulate 5S RNA synthesis above the endogenous levels. Both homologous oocyte RNA polymerase I and III and E. coli RNA polymerase transcribed the 5S genes in deproteinized DNA to approximately the same extent (severalfold above random) and both the sense and anti-sense strands of the gene were transcribed. It appears, therefore, that both chromatin-associated components and a purified RNA polymerase III are necessary and sufficient for the selective and accurate transcription of the 5S RNA genes in vitro.
n-Type Si-based photoelectrochemical cell: New liquid junction photocell using a nonaqueous ferricenium/ferrocene electrolyte
Authors: Legg Kenneth D. , Ellis Arthur B. , Bolts Jeffrey M. , Wrighton Mark S.
Published: May 2006

n-Type Si has been shown to serve as a stable photoanode in a cell for the conversion of light to electricity. The other components of the cell are a Pt cathode and an electrolyte consisting of an ethanol solution of [
n
-Bu
4
N]ClO
4
with a redox couple of ferricenium/ferrocene. Data from a two-compartment cell show that ferrocene is oxidized to ferricenium with 100 ± 2% current efficiency at the Si photoanode. Furthermore, prolonged irradiation of the Si in a one-compartment cell yields constant photocurrent and output characteristics. The maximum open-circuit photopotential is ∼700 mV, and the short-circuit quantum yield for electron flow at low light intensity exceeds 0.5. Conversion of monochromatic 632.8-nm light to electricity with ∼2% power efficiency at an output voltage of ∼200 mV has been sustained. These results represent a stable n-type Si-based photoelectrochemical cell.
Location of protein(s) involved in oligomycin-induced inhibition of mitochondrial adenosinetriphosphatase near the outer surface of the inner membrane
Authors: Maïrouch Hammou , Godinot Catherine
Published: May 2006

Mitoplasts, that is, mitochondria freed from their outer membranes, were prepared from pig heart. Sonication induced an inversion of these mitoplasts, giving inside-out vesicles. Added cytochrome
c
can be bound much better to mitoplasts than to sonicated vesicles; addition of trypsin increased adenosinetriphosphatase (ATPase) (ATP phosphohydrolase; EC 3.6.1.3) activity of sonicated vesicles without significantly affecting that of the mitoplasts. Since the site of fixation of cytochrome
c
was located on the outer side of the inner mitochondrial membrane and since the protein inhibitor of the mitochondrial ATPase is present on the inner face of the inner membrane and is very sensitive to trypsin, it can be concluded that mitoplasts are mainly oriented as normal mitochondria while sonicated vesicles are mainly inverted.

Trypsin treatment can abolish the oligomycin sensitivity of ATPase activity of either mitoplasts or sonicated vesicles. However, trypsin induced the solubilization of the soluble F
1
-ATPase of sonicated vesicles while the ATPase activity remained with the mitoplasts after trypsin action. Therefore, trypsin destroyed the oligomycin effect by rupturing the liaison between F
1
and the membrane in sonicated vesicles. On the other hand, the effect of trypsin on mitoplasts must be attributed to the hydrolysis of a protein located near the outer surface of the inner membrane that is at least structurally involved in the oligomycin sensitivity of the ATPase complex.
A light-activated GTPase in vertebrate photoreceptors: Regulation of light-activated cyclic GMP phosphodiesterase
Authors: Wheeler George L. , Bitensky Mark W.
Published: May 2006

We have been studying the mechanism by which light and nucleoside triphosphates activate the discmembrane phosphodiesterase (oligonucleate 5′-nucleotidohydrolase; EC 3.1.4.1) in frog rod outer segments. GTP is orders of magnitude more effective than ATP as a cofactor in the light-dependent activation step. GTP and the analogue guanylyl-imidodiphosphate function equally as allosteric activators of photoreceptor phosphodiesterase rather than participating in the formation of a phosphorylated activator. Moreover, we have found a light-activated (5-fold) GTPase which participates in the modulation of photoreceptor phosphodiesterase. This GTPase activity appears necessary for the reversal of phosphodiesterase activation
in vitro
and may play a critical role in the
in vivo
regulation of light-sensitive phosphodiesterase. The
K
m
for GTP in the light-activated GTPase reaction is <1 μM. The light sensitivity of this GTPase (number of photons required for half-maximal activation) is identical to that of light-activated phosphodiesterase. The GTPase action spectrum corresponds to the absorption spectrum of rhodopsin. There is, in addition, a light-insensitive GTPase activity with a
K
m
for GTP of 90 μM. At GTP concentrations above 5 μM, there is no appreciable activation of GTPase activity by light. The substrate
K
m
values for guanylate cyclase, light-activated GTPase, and light-activated phosphodiesterase order an enzyme array that might permit light to simultaneously cause the hydrolysis of both the substrate and product of guanylate cyclase. These findings reveal yet another facet of light regulation of photoreceptor/cyclic GMP levels and also provide a striking analogy to the GTP regulation of nonphotoreceptor, hormone-sensitive adenylate cyclase.
Regulation of growth hormone gene expression: synergistic effects of thyroid and glucocorticoid hormones.
Authors: Martial J A , Seeburg P H , Guenzi D , Goodman H M , Baxter J D
Published: May 2006

Cultured rat pituitary cells (GC) respond to thyroid and glucocorticoid hormones by increases in growth hormone production and growth hormone mRNA. When these cells are transferred from medium containing normal animal serum (with 1.8 mug of thyroxine per dl) to a medium containing serum from a thyroidectomized calf, "hypothyroid medium" (with no detectable thyroid hormone), growth hormone production decreases markedly. In cells maintained for 5 days in hypothyroid medium, triiodothyronine induces within 50 hr a 17-fold increase in growth hormone production whereas glucocorticoids, during the same time, produce a negligible (3-fold or less) stimulation. In combination, the two hormones promote a 45-fold stimulation. In all instances the changes in growth hormone production are paralleled by changes in the levels of growth hormone mRNA as measured by cell-free translation. The transfer to hypothyroid medium and the hormonal induction do not affect the relative activities of other mRNAs whose products are detectable on polyacrylamide gels. These studies indicate that thyroid hormone can be an activator of the expression of the growth hormone gene. The results also show that triiodothyronine controls the magnitude of the effect of glucocorticoids on growth hormone mRNA, and provide a model for "permissive" triiodothyronine action. The synergistic effect of these two classes of hormone suggests that they increase levels of growth hormone mRNA by different mechanisms.
Adaptive response due to changes in gene regulation: a study with Drosophila.
Authors: McDonald J F , Chambers G K , David J , Ayala F J
Published: May 2006

In spite of the critical role of the process of adaptation in evolution, there are few detailed studies of the genotypic and molecular basis of the process. Drosophila melanogaster flies selected for increased tolerance to ethanol exhibited higher levels of alcohol dehydrogenase (alcohol:NAD+ oxidoreductase; EC 1.1.1.1) activity than unselected controls. A series of tests (electrophoresis, product inhibition, temperature stability, pH optima, substrate specificity, and Michaelis constants) gave no evidence of structural differences in the enzyme of the selected and the control flies. However, quantitative immunological assays showed that the selected flies contained significantly higher amounts of alcohol dehydrogenase. Adaptation of the selected flies to higher alcohol tolerance has most likely taken place by changes not in the structural gene locus coding for the enzyme, but by regulatory changes affecting the amount of gene product.
Virus-specific proteins in the plasma membrane of cells lytically infected or transformed by pol-oma virus.
Authors: Ito Y , Brocklehurst J R , Dulbecco R
Published: May 2006

Antisera, raised in rats, containing specificities directed against tumor antigen of polyoma virus also react with several proteins present in the plasma membrane of mouse cells infected with the virus. The main component has an apparent molecular weight of 55,000. The appearance of this protein after infection with early temperature-sensitive A mutants was temperature-dependent like tumor antigen itself. Pulse and chase isotope experiments suggest that this protein originates from a precursor, perhaps by cleavage; its production appears to be facilitated by the A mutation. Two other components with apparent molecular weights of 61,000 and 28,000 were also present but were more variable from experiment to experiment. All proteins were absent from the plasma membranes of cells infected with a transformation-defective mutant, NG-18. Up to four virus-specific proteins could be isolated from the plasma membranes of rat, hamster, and mouse cells transformed by the virus. The possible role of the plasma membrane proteins in cell transformation is discussed.
Genes transcribed at diverse rates have a similar conformation in chromatin
Authors: Garel Annie , Zolan Miriam , Axel Richard
Published: May 2006

We have analyzed the DNA generated upon treatment of oviduct nuclei with pancreatic DNase I (deoxyribonucleate 3′-oligonucleotidohydrolase; EC 3.1.4.6), with cDNA copies of specific mRNA sequences to study the structure and organization of transcriptionally active genes in chromatin. In this report we examine the kinetics of digestion of three classes of genes in the oviduct which are transcribed at significantly different rates. Our results indicate that the ovalbumin genes appear to be organized by chromatin proteins in such a way that they are rendered exceedingly sensitive to digestion by DNase I. This sensitivity is not observed in the liver, a tissue in which these genes are transcriptionally inert. Furthermore, the transcriptionally inactive globin genes in the oviduct are not selectively sensitive to nuclease attack and are digested 5 times more slowly in the ovalbumin genes in this tissue. In addition, we have examined the accessibility of a complex subset of genes that are rarely represented in the mRNA and are likely to be transcribed at a frequency orders of magnitude below that of the ovalbumin gene. Comparison of the accessibility of these sequences with that of the ovalbumin gene indicates that these two subsets of genes are recognized and cleaved by DNase I at similar rates. These results suggest that the maintenance of an active conformation about specific genes does not reflect the polymerase distribution about these genes. This active conformation is therefore not confined to sequences actively engaged in the transcription process and may reflect the structure about a subpopulation of the genome which represents the transcriptional potential of a given cell type.
Selective recognition of nucleic acids by proteins: the specificity of guanine interaction with carboxylate ions.
Authors: Lancelot G , Hélène C
Published: May 2006

The interaction of carboxylate ions (acetate, butyrate) with nucleic aicd bases and nucleosides has been investigated by proton magnetic resonance in dimethyl sulfoxide (d6)/H2O mixtures. Carboxylate ions interacted only with guanine derivatives and led to a large downfield shift of the NH2 resonance. A 1:1 stoichiometry was deduced from a study of the concentration dependence of chemical shifts. A study of substituted guanine showed that hydrogen bonding involved N(1)H and NH2(2). An association constant of 110 M-1 was determined. This value is about 30 times higher than the association constant for guanine-cytosine base pair formation under the same experimental conditions. As a matter of fact, carboxylate ions induced a dissociation of guanine-cytosine base pairs. This guanine-carboxylate association is experimental evidence for a highly specific interaction that could play an important role in protein/nucleic acid recognition.
F1-ATPase-catalyzed synthesis of ATP from oleoylphosphate and ADP.
Authors: Johnston R , Criddle R S
Published: May 2006

Purified preparations of F1-ATPase (ATP phosphohydrolase; EC 3.6.1.3) isolated from yeast mitochondria catalyze the reaction of oleoylphosphate with ADP to yield ATP and oleic acid. Formation of ATP is specifically inhibited by the F1-ATPase inhibitor 1799 and by dinitrophenol. In the presence of F1, dinitrophenol "uncouples" the synthase reaction by causing rapid hydrolysis of oleoylphosphate without ATP formation. It is proposed that this F1 catalyzed ATP synthesis reaction corresponds to the terminal chemical step in oxidative phosphorylation.
Sequence arrangement of the 5' ends of simian virus 40 16S and 19S mRNAs.
Authors: Hsu M T , Ford J
Published: May 2006

Electron microscopic examination of molecular hybrids between simian virus 40 DNA, that had been cleaved with EcoRI and then digested with exonuclease III, and either 16S or 19S mRNA produced late during the viral infection cycle indicated that each of these mRNAs contained a 5'-terminal "leader" sequence that was encoded in the viral genome at about map position 0.71-0.75. Hybridization of each of these mRNAs to viral DNA immobilized on nitrocellulose filters supported the electron microscopic observations.
Is evolution gradual or rectangular? Evidence from living fishes.
Authors: Avise J C
Published: May 2006

The traditional view that most evolutionary change is gradual and cumulative within lineages (phyletic gradualism) has recently been challenged by the proposition that the majority of evolutionary change is concentrated within speciation events (rectangular evolution). The logical implications of these competing hypotheses for the means and variances of genetic distance among living members of rapidly and slowly speciating phylads are examined. An example of a critical test of gradual versus rectangular evolution is provided by electrophoretic analyses of genic composition in 69 species of North American Cyprinidae (minnows), and 19 species of Centrarchidae (sunfish). Rate of protein evolution appears somewhat decelerated, if anything, in the rapidly speciating minnows. Results are inconsistent with predictions of rectangular evolution, but are not demonstrably incongruent with predictions of phyletic gradualism.
Chemical characterization of murine Ia alloantigens determined by the i-E/i-C subregions of the H-2 complex.
Authors: Silver J , Russell W A , Reis B L , Frelinger J A
Published: May 2006

Alloantisera directed against the alloantigens determined by the I-E and I-C subregions of the murine major histocompatibility complex precipitate two components that have molecular weights of 35,000 and 29,000. These components, when analyzed by partial NH2-terminal sequencing, show no homology to two components of similar size determined by the I-A subregion. However, the large chain determined by the murine I-E and/or I-C subregion is homologous to the large chain of the human HLA-D region alloantigen, although the small chains isolated from these two species do not display any such homology.
Orientation of bacteriorhodopsin in
Halobacterium halobium
as studied by selective proteolysis
Authors: Gerber Gerhard E. , Gray Christopher P. , Wildenauer Dieter , Khorana H. Gobind
Published: May 2006

The orientation of bacteriorhodopsin in the purple membrane of
Halobacterium halobium
has been studied by proteolytic degradation of purple membrane sheets, reconstituted vesicles, and whole cells, with the following results: (
i
) Bacteriorhodopsin in purple membrane sheets is cleaved at a single site by Pronase or trypsin; a polypeptide segment of about 15 amino acids is lost from the carboxyl end. Carboxypeptidase A sequentially releases amino acids from the carboxyl end; the tetrapeptide sequence -Ala-Ala-Thr-Ser(COOH) was tentatively deduced for this terminus. (
ii
) The apomembrane, which lacks retinal, undergoes a second cleavage with trypsin releasing a fragment of approximately 6300 molecular weight from the amino terminus. (
iii
) Vesicles reconstituted from the purple membrane sheets and synthetic lecithins, in which the direction of proton pumping is opposite to that in the whole cells, have the carboxyl terminus of bacteriorhodopsin accessible to proteolysis. (
iv
) In envelope vesicles, which largely pump protons in the same direction as the whole cells, the carboxyl terminus is largely protected against proteolysis. (
v
) Treatment of whole cells with proteinase K hydrolyzes the cell wall proteins but has no effect on acteriorhodopsin. However, the same treatment after lysis of the cells results in degradation of the hydrophilic region at the carboxyl terminus. The results show that the carboxyl terminus as well as the additional cleavage site near the amino terminus observed in apomembrane are on the cytoplasmic side of the purple membrane.
Oligopeptides as potential antiaggregation agents for deoxyhemoglobin S.
Authors: Kubota S , Yang J T
Published: May 2006

Oligopeptides that mimic segments of the amino acid sequence of hemoglobin S at potential contact sites can be used to inhibit aggregation. These oligopeptide inhibitors raise the minimum gelling concentration of deoxyhemoglobin S so that chemical modification does not have to be used. The hexapeptide amides of both betaS 1-6 which is believed to be one of the contact areas among aggregates, and betaA 1-6 of hemoglobin A increase the minimum gelling concentration by more than 70%. The hexapeptide amide beta79-84 behaves like beta1-6 (beta being betaS or betaA). Shorter oligopeptides, such as betaS3-6, are less effective as an inhibitor but longer ones, such as betaS1-8, are no more effective than beta1-6. Permutations of the sequence, such as betaS125634, do not alter the percent increase in the minimum gelling concentration. Leu- and Met-enkephalin increase the minimum gelling concentration just as beta1-6 does, but (Pro)6 is not very effective. Thus, the use of complementary oligopeptides as inhibitors is extended to include certain "flexible" peptides, which can adapt themselves to interfere with the molecular contacts and thereby gelation of deoxyhemoglobin S.
31P nuclear magnetic resonance measurements of ATPase kinetics in aerobic Escherichia coli cells.
Authors: Brown T R , Ugurbil K , Shulman R G
Published: May 2006

We have measured the in vivo unidirectional rates between the terminal phosphate of ATP and intracellular inorganic phosphate (PiIN) in aerobic suspensions of Escherichia coli cells using 31P nuclear magnetic resonance saturation transfer techniques. Typically, the measurements consisted of saturating the ATPgamma resonance and observing a 20 +/- 5% reduction in the intensity of the PiIN resonance. No saturation transfer was observed after incubation with 1 mM dicyclohexylcarbodiimide, an ATPase inhibitor. From the measured decrease in intensity of PiIN coming from saturation transfer, the apparent unimolecular rate constant of PiIN to ATP was calculated to be 0.6 +/- 0.15 sec-1.
Lymphospecific toxicity in adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency: Possible role of nucleoside kinase(s)
Authors: Carson Dennis A. , Kaye Jonathan , Seegmiller J. E.
Published: May 2006

Inherited deficiencies of the enzymes adenosine deaminase (adenosine aminohydrolase; EC 3.5.4.4) and purine nucleoside phosphorylase (purine-nucleoside:orthophosphate ribosyltransferase; EC 2.4.2.1) preferentially interfere with lymphocyte development while sparing most other organ systems. Previous experiments have shown that through the action of specific kinases, nucleosides can be “trapped” intracellularly in the form of 5′-phosphates. We therefore measured the ability of newborn human tissues to phosphorylate adenosine and deoxyadenosine, the substrate of adenosine deaminase, and also inosine, deoxyinosine, guanosine, and deoxyguanosine, the substrates of purine nucleoside phosphorylase. Substantial activities of adenosine kinase were found in all tissues studied, while guanosine and inosine kinases were detected in none. However, the ability to phosphorylate deoxyadenosine, deoxyinosine, and deoxyguanosine was largely confined to lymphocytes. Adenosine deaminase, but not purine nucleoside phosphorylase, showed a similar lymphoid predominance. Other experiments showed that deoxyadenosine, deoxyinosine, and deoxyguanosine were toxic to human lymphoid cells. The toxicity of deoxyadenosine was reversed by the addition of deoxycytidine, but not uridine, to the culture medium. Based upon these and other experiments, we propose that in adenosine deaminase and purine nucleoside phosphorylase deficiency, toxic deoxyribonucleosides produced by many tissues are selectively trapped in lymphocytes by phosphorylating enzyme(s).
Demonstration of different contractile mechanisms for angiotensin II and des-Asp1-angiotensin II in rabbit aortic strips.
Authors: Ackerly J A , Moore A F , Peach M J
Published: May 2006

Evidence of selective inhibition, differences in dose-response relationships, and cross-tachyphylaxis studies suggest that separate receptors and/or mechanisms may be involved in responses to angiotensin (Ang), [Sar1]Ang II, and Ang III (= des-Asp1-Ang II). The extracellular Ca2+ requirement for contractile responses induced by angiotensin peptides and norepinephrine was determined in rabbit aortic strips. Responses to K+ and [Sar1]Ang II were attenuated markedly by treatment with SKF-525A, verapamil, or Ca2+-free buffer. The response to Ang II was not impaired by verapamil, was blocked partially by SKF-525A, and was reduced markedly in Ca2+-free medium. Norepinephrine- and Ang III-induced contractions were not dependent on extracellular Ca2+. K+, Ang II, and [Sar1]Ang II required extracellular Ca2+ to induce contraction of the rabbit aorta. The data indicate that Ang III may have a mechanism of action that differs from that of [Sar1]Ang II and Ang II.
Mapping of human chromosomal regions related to neoplasia: Evidence from chromosomes 1 and 17
Authors: Rowley Janet D.
Published: May 2006

In clonal aberrations leading to an excess or partial excess of chromosome 1, trisomy for bands 1q25-1q32 was noted in the myeloid cells from all of 34 patients who had various disorders such as acute leukemia, polycythemia vera, and myelofibrosis. This was not the result of a particularly fragile site in that region of the chromosome because the break points in reciprocal translocations that involve it occurred almost exclusively in the short arm. Two consistent rearrangements that have been observed in chromosome 17 produced either duplication of the entire long arm or a translocation of the distal portion of the long arm to chromosome 15. The nonrandom chromosomal changes found in hematologic disorders can now be correlated with the gene loci on these chromosomes or chromosomal segments. Seventy-five genes related to various metabolic enzymes have been mapped; it may be significant that chromosomes carrying gene loci related to nucleic acid metabolism are more frequently involved in hematologic disorders (and other malignancies as well) than are gene loci related to intermediary or carbohydrate metabolism. Furthermore, the known virus-human chromosome associations are closely correlated with the chromosomes affected in hematologic disorders. If one of the effects of carcinogens (including viruses) is to activate genes that regulate host cell DNA synthesis, and if translocations or duplications of specific chromosomal segments produce the same effect, then either of these mechanisms might provide the affected cell with a proliferative advantage.
Opiate receptor: cooperativity of binding observed in brain slices.
Authors: Davis M E , Akera T , Brody T M , Watson L
Published: May 2006

Kinetic constants for binding of 3H-labeled morphine and naloxone were determined from Hill plot analyses and from experiments in which the concentration of tritiated drug was constant and that of nonlabeled drug varied. With brain slices, the binding of either drug exhibited strong positive cooperativity (Hill slope greater than 3); this was not observed in brain homogenates. Thus, in slices the relationship between opiate binding site and ligand may be more relevant physiologically and pharmacologically.
On the equivalence between the boundedness of certain classes of maximal and multiplier operators in Fourier analysis
Authors: Cordoba A. , Fefferman R.
Published: May 2006

The relationship between certain multipliers generalizing the Hilbert transform and maximal operators generalizing the Hardy-Littlewood maximal function is studied and some consequences of this relation are derived.
MacMahon's conjecture on symmetric plane partitions
Authors: Andrews George E.
Published: May 2006

In 1898, P. A. MacMahon conjectured that the generating function for symmetric plane partitions with at most
s
rows and each part at most
j
has an especially simple closed form. An outline of the proof of this conjecture is given here.