bluuebunny/crossref_metadata_2025 huggingface dataset:
Classical geometric resolution of the Einstein—Podolsky—Rosen paradox
Authors: Ne'eman Yuval
Published: May 2006

I show that, in the geometry of a fiber bundle describing a gauge theory, curvature and parallel transport ensure and impose nonseparability. The “Einstein—Podolsky—Rosen paradox” is thus resolved “classically.” I conjecture that the ostentatiously “implausible” features of the quantum treatment are due to the fact that space—time separability, a basic assumption of single-particle nonrelativistic quantum mechanics, does not fit the bundle geometry of the complete physics.
A frameshift mutation affecting the carboxyl terminus of the simian virus 40 large tumor antigen results in a replication- and transformation-defective virus.
Authors: Lewis E D , Chen S , Kumar A , Blanck G , Pollack R E , Manley J L
Published: May 2006

We have constructed a frameshift mutation in the simian virus 40 early region using a novel method of oligonucleotide-directed mutagenesis. The mutated DNA specifies an 84,000-dalton large tumor antigen that consists of approximately equal to 75,000 daltons encoded by the wild-type reading frame and 9,000 daltons, by the alternative reading frame (wild-type large tumor antigen is approximately equal to 82,000 daltons). The frameshifted carboxyl terminus of the protein bears a strong similarity to the same region of polyoma virus middle-sized tumor antigen. We have found that the mutant DNA is unable to replicate when introduced into permissive monkey cells and incapable of transforming nonpermissive mouse cells.
Antibodies to the Epstein-Barr virus nuclear antigen and to rheumatoid arthritis nuclear antigen identify the same polypeptide.
Authors: Billings P B , Hoch S O , White P J , Carson D A , Vaughan J H
Published: May 2006

Patients with seropositive rheumatoid arthritis (RA) have elevated titers of precipitating antibody toward an antigen designated RA nuclear antigen (RANA). Anti-RANA reactivity has been associated with prior Epstein-Barr virus (EBV) infection. Using the protein blot technique, we have identified, in extracts of WI-L2, an EBV+ nonproducer B-lymphoblast line, a Mr 80,000 polypeptide that is reactive with anti-RANA-containing sera. This same polypeptide can be recovered from RANA precipitin bands. The Mr 80,000 polypeptide appears to be EBV-associated, as a homologous antigen was detected in two other EBV+ cell lines, Daudi and Raji, but was not present in three EBV- human cell lines tested, HeLa, BJAB, and Ramos. Anti-RANA antibodies and antibodies reactive with the Mr 80,000 polypeptide also appear coincidently in the sera of individuals exhibiting an EBV infection (infectious mononucleosis). Further analysis of the RANA-associated Mr 80,000 polypeptide suggested its identity with the previously recognized EBV-associated nuclear antigen designated EBNA. The Mr 80,000 antigen shares with EBNA the properties of being a heat-stable, DNA binding protein. EBNA is traditionally assayed by a complement fixation reaction and anti-Mr 80,000 antibodies were shown to be reactive when a complement fixation assay was used in the immunoblot. Finally, when the Mr 80,000 antigen was used to absorb an anti-RANA/anti-EBNA serum, both antibodies were reduced.
Identification of a cytoplasmic tropomyosin gene linked to two muscle tropomyosin genes in Drosophila.
Authors: Bautch V L , Storti R V
Published: May 2006

A Drosophila cytoplasmic tropomyosin gene has been identified and is located on the same genomic DNA clone as two Drosophila muscle tropomyosin genes previously identified. A positive hybrid-selection translation assay using the subcloned gene and RNA from non-muscle cell sources yielded a protein with a size (Mr, 31,000) and isoelectric point (5.0) similar to vertebrate cytoplasmic tropomyosin. A modified protocol for the purification of vertebrate cytoplasmic tropomyosin was used to partially purify cytoplasmic tropomyosin from the Drosophila Kc cell line. The Kc cell protein was identified as a cytoplasmic form of tropomyosin on the basis of its size, isoelectric point, and crossreactivity with a polyclonal vertebrate antitropomyosin antibody in a two-step binding assay. The Kc cell cytoplasmic tropomyosin comigrates in two dimensions with the hybrid-selected in vitro translation product of the region 3 gene, and both proteins show a mobility shift in NaDodSO4/urea/polyacrylamide gels that is characteristic of vertebrate tropomyosins. The cytoplasmic tropomyosin gene hybridizes to both Drosophila muscle tropomyosin genes under decreased stringency conditions. This cross-hybridization spans several internal restriction endonuclease sites in each muscle tropomyosin gene and indicates an overall partial homology among the three Drosophila tropomyosin genes. These results show that Drosophila tropomyosins are encoded by a closely linked family of differentially regulated genes.
Point mutations and DNA rearrangements 5' to the inducible qa-2 gene of Neurospora allow activator protein-independent transcription.
Authors: Geever R F , Case M E , Tyler B M , Buxton F , Giles N H
Published: May 2006

Expression of the qa-2 gene of Neurospora crassa normally requires a functional activator protein encoded by qa-1F. Twelve transcriptional mutants of the qa-2 gene have been isolated in qa-1F- strains, and these allow partial expression of qa-2 (1-45% of induced wild type) in the absence of functional activator protein. All 12 mutants have been characterized by genomic (Southern) blot hybridization and the DNAs of 5 have been cloned and sequenced. Eight mutations consist of large DNA rearrangements within a 500-base-pair region 5' to the qa-2 gene. One large rearrangement mutation, located 378 base pairs before the normal site of transcription initiation, causes exceptional levels of qa-2 transcription (45% of induced wild type) from near the normal initiation site. Two of the other four mutations cloned involve tandem duplications (68 and 84 base pairs) of the same upstream region (centered at nucleotide - 145), and two involve "point" mutations (at nucleotides -200 and -95) that closely flank the duplicated region. With one possible exception, none of the mutations appears to involve changes directly associated with RNA polymerase II binding and hence they differ from analogous mutations in comparable prokaryotic systems. The overall results suggest that at least some of the large DNA rearrangement mutations may be acting as upstream activator elements, possibly by juxtaposing enhancer-like sequences, whereas the duplications and point mutations may define a region of qa-2 regulation, for instance at the level of RNA polymerase II access.
Stopped-flow cryoenzymology: detection of catalytic intermediates not observable at ambient temperatures.
Authors: Van Wart H E , Lin S H
Published: May 2006

Stopped-flow cryoenzymology has been used to study the reaction of porcine kidney cytosol leucine aminopeptidase [alpha-aminoacyl-peptide hydrolase(cytosol), EC 3.4.11.1] with L-leucylglycyldansyl hydrazide in 50% (vol/vol) methanol buffer over the -40 to 23 degrees C temperature range. Resonance energy transfer between tryptophan residues of the enzyme E and the dansyl group of the substrate S has been used to detect the formation and interconversion of reaction intermediates (E X S)i. Above 0 degrees C, a single intermediate E X S is formed and decays by first-order kinetics to products. However, at temperatures below -20 degrees C, a new intermediate (E X S)' is observed immediately after mixing, which relaxes to E X S within 100 msec. Because the detection of this new intermediate would not have been possible at ambient temperatures, this illustrates the value of stopped-flow cryoenzymology for studies of catalytic pathways.
Enhancement of spontaneous mitotic recombination by the meiotic mutant spo11-1 in Saccharomyces cerevisiae.
Authors: Bruschi C V , Esposito M S
Published: May 2006

Both nonreciprocal and reciprocal mitotic recombination are enhanced by the recessive mutant spo11-1, which was previously shown to affect meiosis by decreasing recombination and increasing nondisjunction. The mitotic effects are not distributed equally in all chromosomal regions. The genotypes of mitotic recombinants in spo11-1/spo11-1 diploid cells provide further evidence that widely spaced chromosomal markers undergo coincident conversion in mitosis.
Quantitative correlation of homogeneously stained regions on chromosome 10 with dihydrofolate reductase enzyme in human cells.
Authors: Wolman S R , Craven M L , Grill S P , Domin B A , Cheng Y C
Published: May 2006

Gene amplification may be visualized within a chromosome as a homogeneously stained region (HSR) and HSRs have rarely been reported in human tumor cells with identification of the amplified gene. A parental line and seven clones derived from KB cells resistant to methotrexate (MTX) contain dihydrofolate reductase (DHFR; tetrahydrofolate dehydrogenase; EC 1.5.1.3), ranging from 0.007 unit/mg in the parent to 0.369 unit/mg in clone 7A with a 13,000-fold increase in resistance to MTX. The enzyme is identical to DHFR from other human sources, including that from leukemic patients. A HSR localized to the long arm of chromosome 10(q26) is present in clones selected at or above 2.5 microM MTX. Increase in number of 10q per cell, increase in number of HSR, and increasing amounts of DHFR correlate well. The chromosome change is stable with time as is enzyme production even in the absence of selection by MTX. No clone has shown double minutes. The gene copy number is low. The stability and low gene copy in the presence of large HSRs differ from the pattern described for murine tumors. A human gene for DHFR may be associated with the long arm of chromosome 10.
High-frequency mutation at the adenine phosphoribosyltransferase locus in Chinese hamster ovary cells due to deletion of the gene.
Authors: Simon A E , Taylor M W
Published: May 2006

Evidence for a two-step model to explain the high-frequency expression of the recessive phenotype at the autosomal adenine phosphoribosyltransferase (APRT; EC 2.4.2.7) locus in Chinese hamster ovary (CHO) cells was given by Simon et al. [Simon, A. E., Taylor, M. W., Bradley, W. E. C. & Thompson, L. (1982) Mol. Cell. Biol. 2, 1126-1133]. This model proposed a high-frequency event, leading to allelic inactivation or a loss of gene function, and a low-frequency event, causing a structural alteration of the APRT protein. Either event could occur first, resulting in two classes of heterozygotes. We have analyzed the low-frequency event that gave rise to the class 2 aprt heterozygote D416 and the high-frequency event that led to APRT- cells derived from D416. Genomic Southern blots of Msp I- or Hpa II-digested DNA from wild-type CHO, aprt heterozygote D416, and two APRT- cell lines derived from D416 indicate a loss of a specific Msp I/Hpa II recognition sequence at one aprt locus in the heterozygote that correlates with the production of the electrophoretically altered APRT protein found in D416. The APRT- mutants are homozygous for the loss of this Msp I/Hpa II site. By using an additional CHO gene as an internal control, it was determined that the APRT- mutants contain only a single copy of the altered aprt gene. Thus, the high-frequency event that produces APRT- mutants derived from D416 is not an inactivation event but rather a deletion of the wild-type aprt gene.
Partial disruption of naturally occurring groups of insulin receptors on adipocyte plasma membranes by dithiothreitol and N-ethylmaleimide: the role of disulfide bonds.
Authors: Jarett L , Smith R M
Published: May 2006

In this ultrastructural study, monomeric ferritin-insulin was used to further elucidate the role of disulfide bonds in maintaining the natural groups of insulin receptors on adipocyte plasma membranes. Dithiothreitol (1 mM) caused partial disruption of the occupied receptor groups with an increase in single receptors to greater than 50% of total occupied receptors. N-Ethylmaleimide (1 mM) disrupted the groups to the same extent as dithiothreitol and the effect was partly additive with the dithiothreitol effect. The magnitude of the disruption caused by dithiothreitol or N-ethylmaleimide was similar to that caused by cytochalasin B. Dithiothreitol, a reducing agent, caused a marked increase in binding of insulin to the plasma membranes while N-ethylmaleimide and cytochalasin B, both thiol reagents, had little if any effect on insulin binding. These data suggest that two different sets of disulfide bonds are involved. One set was susceptible to both reducing and thiol reagents and responsible for holding the receptor groups together, and the other set was susceptible to reducing agents only and related to the increased insulin binding caused by dithiothreitol. A proposed model is discussed.
Construction of a shuttle vector for the filamentous fungus Neurospora crassa.
Authors: Stohl L L , Lambowitz A M
Published: May 2006

We have constructed a recombinant plasmid, pALS-1, that replicates autonomously in both Neurospora and Escherichia coli. pALS-1 consists of the mitochondrial plasmid from Neurospora strain P405-Labelle, the Neurospora qa-2+ gene, and E. coli plasmid pBR325. pALS-1 transforms the Neurospora qa-2+ gene at frequencies 5- to 10-fold higher than those for plasmids that transform mainly by integration. When E. coli was transformed with DNA from Neurospora transformants, we recovered not only pALS-1 but also a smaller plasmid, pALS-2, which had undergone deletion of most and possibly all Labelle sequences, and the immediately flanking sequences in pBR325. pALS-2 also appears to replicate autonomously in Neurospora, but less efficiently than does pALS-1. Southern blots show that free pALS-1 and pALS-2 are present in nuclear and cytosolic (supernatant from high-speed centrifugation) fractions of Neurospora transformants and that small, variable proportions of the plasmids also can be detected in mitochondria. pALS-1 and pALS-2 constitute putative shuttle vectors for Neurospora.
DNA sequence changes in an upstream DNase I-hypersensitive region are correlated with reduced gene expression.
Authors: McGinnis W , Shermoen A W , Heemskerk J , Beckendorf S K
Published: May 2006

Previous experiments have identified a region that is required for the expression of the Drosophila glue protein gene Sgs-4 and is located 300-500 base pairs upstream from the structural gene. The chromatin in this region changes conformation and becomes hypersensitive to DNase I digestion when the gene becomes active, a change that apparently induces additional conformational changes near the site of transcription initiation. To learn more about the DNA sequence requirements for the function of this region, we analyzed three naturally occurring Sgs-4 under-producers. In two of these strains, a single base pair change within the hypersensitive region is correlated with a 50% reduction in the amount of Sgs-4 RNA produced. Another strain, which has multiple 5' lesions, is severely reduced in Sgs-4 expression and in the DNase hypersensitivity of the upstream region. Several of the sequence changes in this extreme underproducer lie near hypersensitive sites, suggesting that they inhibit the appearance of the normal DNase hypersensitive conformation.
In vivo expression of rat insulin after intravenous administration of the liposome-entrapped gene for rat insulin I.
Authors: Nicolau C , Le Pape A , Soriano P , Fargette F , Juhel M F
Published: May 2006

A recombinant plasmid encoding rat preproinsulin I was encapsulated in large liposomes and intravenously injected in rats. Glycemia and blood, splenic, and hepatic insulin were assayed at various times beginning 6 hr after inoculation. The results were compared with controls that had received (i) empty liposomes, (ii) liposomes carrying the Escherichia coli pBR322 plasmid, (iii) the free rat insulin I gene, and (iv) no injection at all. Whereas all controls showed unchanged glucose and insulin levels, the treated animals had, 6 hr after inoculation, a blood glucose level of 72 +/- 5 mg of glucose/100 ml of blood as compared with 107 +/- 2 mg/ml for controls. Radioimmunoassay of blood insulin gave 61 +/- 8 microunits/ml as compared with 43 +/- 5 microunits/ml for controls and the spleen and liver values were 242 +/- 22 and 204 +/- 20 microunits/g of tissue, respectively, as compared with 112 +/- 20 and 87 +/- 15 microunits/g in controls. External gamma-camera imaging of the organ uptake of 111In-labeled liposomes permitted study of the kinetics and extent of uptake of the liposomes by spleen and liver, results that support the findings concerning insulin synthesis in the two organs.
Nucleotide and deduced amino acid sequences of Torpedo californica acetylcholine receptor gamma subunit.
Authors: Claudio T , Ballivet M , Patrick J , Heinemann S
Published: May 2006

The nucleotide sequence has been determined of a cDNA clone that codes for the 60,000-dalton gamma subunit of Torpedo californica acetylcholine receptor. The length of the cDNA clone is 2,010 base pairs. The 5' and 3' untranslated regions have respective lengths of 31 and 461 base pairs. Data suggest that the putative polyadenylylation consensus sequence A-A-T-A-A-A may not be required for polyadenylylation of the mRNA corresponding to the cDNA clone described in this study. From the DNA sequence data, the amino acid sequence of the gamma subunit was deduced. The subunit is composed of 489 amino acids giving a molecular mass of 56,600 daltons. The deduced amino acid sequence data also indicate the presence of a 17-amino acid extension or signal peptide on this subunit. From these data, structural predictions for the gamma subunit are made such as potential membrane-spanning regions, possible asparagine-linked glycosylation sites, and the assignment of regions of the protein to the extracellular, internal, and cytoplasmic domains of the lipid bilayer.
Platelet-activating factor stimulates metabolism of phosphoinositides in horse platelets: possible relationship to Ca2+ mobilization during stimulation.
Authors: Billah M M , Lapetina E G
Published: May 2006

Stimulation of horse platelets with platelet-activating factor (PAF) induces a rapid degradation of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. Addition of 0.1 microM PAF for 5 sec to platelets prelabeled with 32P induces a 50% loss of [32P]PtdIns(4,5)P2. 32P-Labeled phosphatidylinositol 4-monophosphate (PtdIns4P) and [32P]phosphatidylinositol (PtdIns) also are decreased, albeit at a slower rate. Loss of 32P radioactivity correlates with a net loss of fatty acids from both polyphosphoinositides. Stimulation of platelets with PAF also produces formation of [32P]phosphatidic acid and [32P]lysophosphatidylinositol. The initial disappearance of inositol lipids is subsequently followed by resynthesis, as evidenced by increased incorporation of 32P into PtdIns(4,5)P2, PtdIns4P, and PtdIns. The resynthesis of the inositides increases with time and is proportional to the concentration of PAF. Prostacyclin (1 microM) inhibits (i) the formation of phosphatidic acid and lysophosphatidylinositol and (ii) the resynthesis of polyphosphoinositides induced by 0.03 microM PAF without affecting the initial loss of PtdIns(4,5)P2. The loss of inositol lipids appears to be a primary event of platelet activation. The initial loss of polyphosphoinositides might be linked to the initiation of cellular activation by mobilizing membrane-bound Ca2+, whereas the subsequent formation of these lipids might be involved in mechanisms to prevent overstimulation of the cell.
Mechanism of action of glycopeptide hormones and cholera toxin: what is the role of ADP-ribosylation?
Authors: Rebois R V , Beckner S K , Brady R O , Fishman P H
Published: May 2006

The cultured murine Leydig tumor cell line MLTC-1 and the normal rat thyroid strain FRTL have adenylate cyclase activities that are stimulated by human chorionic gonadotropin (hCG) and thyrotropin, respectively. Both cell types also respond to choleragen. Activation of adenylate cyclase in membranes by choleragen required NAD whereas stimulation of the enzyme by hormones did not. With [alpha-32P]NAD as a donor, ADP-ribosylation of membrane proteins was determined under the same conditions used to assay adenylate cyclase activity. Under these conditions, choleragen, but not the hormones, caused the ADP-ribosylation of subunits of the regulatory component (G/F) of adenylate cyclase in both FRTL and MLTC-1 membranes. In the absence of any effectors, several membrane proteins became labeled but the hormones did not cause the specific labeling of these or any other membrane proteins. Pretreatment of intact MLTC-1 cells with hCG did not block the ability of choleragen to ADP-ribosylate G/F in isolated membranes; labeling was actually enhanced in a manner related to the length of exposure to hCG. In contrast, pretreatment of the cells with choleragen inhibited ADP-ribosylation of G/F by the toxin in isolated membranes. Extracts of membranes from untreated, hCG-treated, and choleragen-treated MLTC-1 cells were used to reconstitute adenylate cyclase activity in membranes from the cyc- variant of S49 lymphoma cells which lacks a functional G/F. Toxin but not hormone treatment caused an increase in the basal activity of adenylate cyclase in the reconstituted system. Our results indicate that ADP-ribosylation of the regulatory component of adenylate cyclase is required for choleragen action but not for hormone action.
Environmental controls on stomatal conductance in a shrub of the humid tropics
Authors: Mooney H. A. , Field C. , Yanes C. Vásquez , Chu C.
Published: May 2006

Leaves of
Piper hispidum
, a shrub native to the lowland tropics of Mexico, have a strong stomatal response to humidity that results in similar rates of water loss under a wide range of leaf-to-air water-vapor concentration gradients. Stomatal conductance of these leaves is insensitive to CO
2
concentration and increases in response to high humidity even in the dark.
Membrane insertion at the leading edge of motile fibroblasts.
Authors: Bergmann J E , Kupfer A , Singer S J
Published: May 2006

We are concerned with the mechanisms involved in the directed migration of eukaryotic cells. Previously we found that, inside cells at the edge of an experimental wound, the Golgi apparatus and the microtubule-organizing center were rapidly repositioned forward of the nucleus in the direction of subsequent cell migration into the wound. This repositioning was proposed to serve the purpose of introducing new membrane mass at the leading edge of the cell, by directing Golgi apparatus-derived vesicles bound for the plasma membrane to that edge. We now provide evidence to support this proposal. Cultured fibroblastic cells at the edge of a wound were infected with a temperature-sensitive mutant (0-45) of vesicular stomatitis virus. It is known that the G-protein, an integral membrane protein of the virus, is synthesized and remains in the rough endoplasmic reticulum at the nonpermissive temperature, but when the infected cells are shifted to the permissive temperature, the G-protein moves through the Golgi apparatus to the plasma membrane. By immunofluorescence microscopy, we here show that the first appearance of the G-protein at the cell surface corresponds to the leading edge of the motile cell. These observations are incorporated into a coherent scheme for the mechanisms involved in cell migration.
Relation of cell surface antigens on methylcholanthrene-induced fibrosarcomas to immunoglobulin heavy chain complex variable region-linked T cell interaction molecules
Authors: Flood Patrick M. , DeLeo Albert B. , Old Lloyd J. , Gershon Richard K.
Published: May 2006

For optimal activation, T suppressor cells must receive a signal froma specialized inducer cell. The activating signal is delivered by a molecular complex that is composed, in part, of polymorphic gene products that are controlled by the immunoglobulin heavy chain complex (
Igh
) and that map to the variable region (
Igh-V
) of the complex. Consequently, if suppressor/inducer cells and their acceptor cells do not share
Igh-V
-encoded polymorphisms, the inducer molecules fail to activate the suppressor cells. Other polymorphic gene products (those that act as transplantation antigens on methylcholanthrene-induced fibrosarcomas) have been shown to be encoded by a gene(s) that maps to the same general region of chromosome 12 as does the
Igh
complex. Therefore, we tested the hypothesis that polymorphic gene products expressed by the T suppressor/inducer cell and its biologically active product were related to the transplantation antigens on these fibrosarcomas. We have found that isoantisera directed to a number of BALB/c methylcholanthrene-induced tumors block the induction of T suppressor cells but they do so only if the inducer or acceptor T cell (or both) expresses BALB/c
Igh
-linked polymorphisms. All other BALB/c gene products are irrelevant. These findings suggest that “transformation related antigens” on some sarcoma cells are variants of, or are otherwise related to, the cell interaction molecules that T cell subsets use to communicate with one another under normal circumstances. Furthermore, the findings that (
a
) multiple methylcholanthrene-induced tumors express serologically related antigens (as well as the unique ones that have been previously demonstrated by transplantation tests) and (
b
) these serologically related antigens can be found on
Igh-V
-controlled T cell communication molecules raise the intriguing possibility that, like certain viruses, methylcholanthrene reacts with specific regions of cellular DNA that either directly or indirectly regulate the expression of other specific genes (in this particular case the genes that encode the T cell communication molecules).
Rescue of transforming Epstein-Barr virus (EBV) from EBV-genome-positive epithelial hybrid cells transfected with subgenomic fragments of EBV DNA
Authors: Stoerker Jay , Glaser Ronald
Published: May 2006

Transfection experiments using subgenomic fragments of the B95-8 strain of Epstein-Barr virus (EBV) DNA and EBV genome (HR-1)-positive epithelial/Burkitt hybrid cells (D98/HR-1) were carried out to determine whether an interaction between the transfecting virus fragment(s) and the endogenous HR-1 EBV genome could take place. Expression of EBV-specific antigens, including early antigen and virus capsid antigen, was examined in transfected cells by immunofluorescence. Attempts were also made to recover biologically active EBV from the D98/HR-1 cells after transfection with cloned fragments of B95-8 DNA. We found that D98/HR-1 cells transfected with the
Bam
HI H or H, F, and X fragments were positive for early antigen 3 days after transfection. Spent media from transfected D98/HR-1 cells maintained for 20-30 days in culture were pooled, filtered, concentrated, and used as a potential source of virus to inoculate human umbilical cord blood lymphocytes. No evidence of transformation was observed with such preparations. However, if spent medium from D98/HR-1 cell cultures was first treated with iododeoxyuridine (to induce EBV DNA synthesis and replicative cycle) and then transfected with the
Bam
HI H, F, and X fragments of B95-8 DNA and used to infect cord blood lymphocytes, transformation was obtained. A lymphoblastoid cell line derived in this manner, designated HI-HFX, is an EBV nuclear antigen-positive nonproducer cell line. Similar results were obtained with preparations from iododeoxyuridine-treated D98/HR-1 cells transfected with the EB 26-36 fragment of B95-8 DNA cloned in a Charon 4A vector. The EB 26-36 fragment contains the
Bam
HI H, F, and X regions.
Magellanic cloud planetary nebula with suspected strong forbidden iron lines
Authors: Aller L. H. , Czyzak S. J.
Published: May 2006

The relatively high-excitation nebula (Westerlund-Smith object 25) in the large Magellanic cloud shows prominent forbidden lines of [Ar IV], the close [Ne IV] pair λ4724, 4726, [Ca V] λ5309, [Fe V] λ4227, and probably [Fe VI] and [Fe VII], as well. A conventional interpretation of observations secured with a vidicon detector at the Cerro Tololo 4-m telescopes indicates an essentially “normal” helium abundance but depletions of N, O, Ne, and other elements with respect to our own galaxy. When a comparison is made with diffuse nebulae or H II regions in the large Magellanic cloud, we find helium to be more abundant, oxygen to be depleted, and nitrogen, neon, and argon to be comparable. The abundance of sulfur is uncertain. Iron in the gaseous phase is certainly more plentiful than in conventional planetaries.
Histone mRNA concentrations are regulated at the level of transcription and mRNA degradation.
Authors: Sittman D B , Graves R A , Marzluff W F
Published: May 2006

The levels of histone mRNA are rapidly reduced after treatment of cultured cells with hydroxyurea or cytosine arabinonucleoside. The histone mRNA for the replicative histone variants is destroyed rapidly, with a half-life of 10-15 min. The levels of mRNA coding for the replacement histone variant H3.3 were unchanged after treatment with DNA synthesis inhibitors. In addition to the rapid destruction of histone mRNA, there was a reduction to 1/5th in the rate of transcription of the histone genes. Lymphoma cells (S49) arrested in G1 by cyclic AMP produce and contain significant levels of histone mRNA. Hydroxyurea reduces the rate of transcription and the levels of histone mRNA in the G1-arrested cells.
Homologous recombination in mammalian cells mediates formation of a functional gene from two overlapping gene fragments.
Authors: de Saint Vincent B R , Wahl G M
Published: May 2006

Chinese hamster cells with a lesion in the CAD gene (cell line Urd-A) require exogenous uridine to survive. Uridine prototrophs could be isolated after introducing two recombinant plasmids containing overlapping fragments of a cloned Syrian hamster CAD gene. In contrast, no uridine prototrophs were obtained after introducing a plasmid containing only one of the two overlapping fragments. DNA restriction analysis showed that the prototrophic transformants contain a functional CAD gene which was formed by a recombination event in the overlapping region of the two clones. Most of the recombination events involved homologous exchanges, and some of them apparently were reciprocal. In situ hybridization analysis revealed that the donated sequences were integrated at a single chromosomal site which was different in each transformant. These results demonstrate the existence of a recombination system(s) in mammalian cells that can catalyze homologous exchanges. Recombination between donated sequences is a means by which this system can be characterized and also utilized for the production of novel gene fusions.
Alloprocoptic selection: A mode of natural selection promoting polymorphism
Authors: Serradilla Juan M. , Ayala Francisco J.
Published: May 2006

The fitness effect of genetic variation at three loci coding for enzymes is studied in
Drosophila melanogaster
. The fertility of a female is determined by the female genotype as well as by the genotype of the male with which she mates. Significant interactions exist between female and male genotype, so that the fertility of a given mating combination cannot be predicted from the average fertility of the two genotypes involved. Multiple stable equilibria are possible when such interactions exist. At two loci, the fertility is greater than expected when the two mating individuals are homozygous for different alleles and smaller than expected when they are homozygous for the same allele. This mode of selection in which association of opposites increases their fitness is herein named alloprocoptic selection. It will contribute to maintaining genetic polymorphism in nature.
Gonadotropin-releasing hormone (GnRH) stimulates phosphatidylinositol metabolism in rat granulosa cells: mechanism of action of GnRH.
Authors: Davis J S , Farese R V , Clark M R
Published: May 2006

This report describes the effects of gonadotropin-releasing hormone (GnRH; gonadoliberin) and an agonist, [D-Ala6, des-Gly10]GnRH ethyl amide (GnRHa), on phospholipid metabolism in rat granulosa cells isolated from mature Graafian follicles. As indicated by the incorporation of 32PO4, GnRHa rapidly (less than 2 min) stimulated the labeling of phosphatidic acid and phosphatidylinositol but had no effect on the labeling of other phospholipids. Increased phosphatidylinositol labeling was also observed when myo-[2-3H]inositol was incubated with granulosa cells in the presence of GnRHa. Increases in labeling were dependent on the dose of GnRH and time of incubation. Thyrotropin-releasing hormone and a specific GnRH antagonist had no effect on labeling, but a GnRH antagonist prevented the stimulatory action of GnRH. In addition, treatment with GnRHa slightly increased the levels of phosphatidylinositol (15%) in 60-min incubations but had no effect on the levels of other phospholipids. Significant increases in progesterone accumulation were observed after 30 min of incubation with GnRHa, and further increases were correlated with the time of incubation. The stimulatory action of GnRH on phospholipid metabolism and progesterone accumulation was not related to increases in cyclic nucleotide accumulation. In incubations lasting up to 30 min, GnRHa had no effect on cAMP accumulation. However, a transient decrease in cGMP levels was observed in response to GnRHa. These studies suggest that the rapid and specific effects of GnRH on phospholipid metabolism in rat granulosa cells represent early events in the action of GnRH.
Structural and functional evidence for multiple channel complexes in the outer membrane of Neurospora crassa mitochondria.
Authors: Mannella C A , Colombini M , Frank J
Published: May 2006

The outer membrane of mitochondria contains proteins that form channels called VDAC (voltage-dependent anion-selective channels). Two independent lines of evidence suggest that these channels occur in specific complexes in the outer membrane of Neurospora mitochondria. Electron microscopic images of these outer membranes reveal polymorphic crystalline arrays of putative pores. These arrays can be shown to be interrelated by movement in the membrane plane of a particular rigid channel triplet or of regular aggregates of this triplet. Detergent extracts of the same outer membranes induce single- and multiple-step conductances in planar phospholipid membranes, with a marked preference for the insertion of triplets and multiples of triplets. The tendency of the mitochondrial channels to occur as extended arrays, apparently built up from triplets, may have important functional and evolutionary implications.
Rapid changes in specific estrogen binding elicited by cGMP or cAMP in cytosol from human endometrial cells.
Authors: Fleming H , Blumenthal R , Gurpide E
Published: May 2006

Addition of cGMP to cytosol of human endometrium or to cells of the endometrial cancer line HEC-1 produced severalfold increases in specific estrogen binding (EB) levels. This effect was maximal with 1 microM cGMP in the presence of 0.1 mM isobutylmethylxanthine (a phosphodiesterase inhibitor) during incubations with [3H]estradiol. In contrast, cAMP decreased EB levels under similar conditions. The effects of cyclic nucleotides on EB levels were complete in less than 15 min in the presence of Mg2+, Mn2+, or Ca2+. The EB sites generated by the addition of cGMP during labeling of cytosol with 10 nM [3H]estradiol were found to sediment in the 8S and 4S regions of low-salt glycerol gradients. No changes in EB levels were observed when cyclic nucleotides were added to cytosol depleted of ATP by preincubation at 4 degrees C for 3 hr, but responsiveness was restored by addition of exogenous ATP. The ATP requirement and the pattern of dependence of cyclic nucleotide actions on divalent cation concentrations suggest that cGMP and cAMP effects may be mediated by kinases and may involve phosphorylations. Another possibility is that the cyclic nucleotides interact allosterically with the binder in the presence of ATP. Addition of sodium molybdate, ATP, and GTP to homogenates of endometrial tissue or HEC-1 cells produces increases in EB levels similar to those obtained by the addition of cGMP. However, these compounds are much less active when added to cytoplasm or cytosol. On the basis of these and other observations, it is hypothesized that molybdate, ATP, and GTP affect EB levels primarily by increasing cGMP concentrations through processes involving a plasma membrane-bound guanylate cyclase.
Pyrimidine metabolism in Tritrichomonas foetus.
Authors: Wang C C , Verham R , Tzeng S F , Aldritt S , Cheng H W
Published: May 2006

The anaerobic parasitic protozoa Tritrichomonas foetus is found incapable of de novo pyrimidine biosynthesis by its failure to incorporate bicarbonate, aspartate, or orotate into pyrimidine nucleotides or nucleic acids. Uracil phosphoribosyltransferase in the cytoplasm provides the major pyrimidine salvage for the parasite. Exogenous uridine and cytidine are mostly converted to uracil by uridine phosphorylase and cytidine deaminase in T. foetus prior to incorporation. T. foetus cannot incorporate labels from exogenous uracil or uridine into DNA; it has no detectable dihydrofolate reductase or thymidylate synthetase and is resistant to methotrexate, pyrimethamine, trimethoprim, and 5-bromovinyldeoxyuridine at millimolar concentrations. It has an enzyme thymidine phosphotransferase in cellular fraction pelleting at 100,000 X g that can convert exogenous thymidine to TMP via a phosphate donor such as p-nitrophenyl phosphate or nucleoside 5'-monophosphate. Thymidine salvage in T. foetus is thus totally dissociated from other pyrimidine salvage.
Derivation of the relation between the linear Onsager coefficients and the equilibrium one-way cycle fluxes of a biochemical kinetic diagram
Authors: Hill Terrell L.
Published: May 2006

The Onsager flux-force coefficients

L
ij

for any biochemical system at a steady state near equilibrium have a simple interpretation: they are linear combinations of the equilibrium one-way cycle fluxes of the kinetic diagram for the system. An explicit and general expression for

L
ij

is derived. This supplements the more pedagogical argument published elsewhere, which was based on a particular special case.
Rapid degradation of unassembled ribulose 1,5-bisphosphate carboxylase small subunits in chloroplasts
Authors: Schmidt Gregory W. , Mishkind Michael L.
Published: May 2006

We have detected a proteolytic mechanism in chloroplasts that selectively and rapidly degrades the imported small subunit of ribulose 1,5-bisphosphate carboxylase when pools of the chloroplast-synthesized large subunit are depleted. This degradation system is constitutively present and appears to be responsible for precise stoichiometric accumulation of the two subunits of the enzyme. We believe similar proteolytic mechanisms participate in regulating the accumulation of other photosynthetic proteins during chloroplast biogenesis.
Tunneling in
Chromatium
chromatophores: Detection of a Hopfield charge-transfer band
Authors: Goldstein Robert F. , Bearden Alan
Published: May 2006

We have observed a weak charge-transfer band in the cytochrome
c
-P
870
electron-transfer reaction in
Chromatium vinosum
chromatophores at 10 K and at 85 K. First, the intermediate acceptor, I, was trapped in the reduced state by lowering the redox potential at room temperature, then illuminating with white light at low temperature for 20 min. Next, illumination by broadband infrared (1-3 μm, 6.5 kW/m
2
) for 4 hr at 10 K decreased the I
-
electron spin resonance signal by 30%. One-hour infrared illumination at 85 K decreased the cytochrome
c
Soret band shift by 10%. The effect of infrared was to promote the system from the ground vibrational state with the electron on P
870
to an excited vibrational state with the electron on cytochrome
c
. The absorption band peak is near 2 μm, and the integrated cross section is ≈6 × 10
-3
eV·M
-1
·cm
-1
. These values are consistent with small (0.02 nm) nuclear motion and with electron-transfer rates measured in the dark.
Mapping of the late promoter of simian virus 40.
Authors: Hartzell S W , Byrne B J , Subramanian K N
Published: May 2006

Mapping of the simian virus 40 (SV40) late promoter was carried out in the absence of the viral early protein, large tumor (T) antigen, and replication of the viral DNA template. SV40 late control region DNA fragments, containing specific deletions, were cloned directly upstream from the coding region of the herpes simplex virus-1 (HSV-1) thymidine kinase (TK; ATP:thymidine 5'-phosphotransferase, EC 2.7.1.21) gene (tk). The promoter activities of the fragments were determined by measuring the tk transformation frequencies of the chimeric tk constructs in mouse L TK- APRT- (adenine phosphoribosyltransferase-negative) and human 143B TK- cell lines. The following results were obtained. (i) The SV40 control region functions with equal efficiencies in the early and late promoter orientations. (ii) A major late control element has been localized within the G+C-rich 21-base-pair (bp) repeat. Thus, in conjunction with our earlier results, the 21-bp repeat is a bidirectional promoter element functioning as a major component of both the early and late promoters and is an element that enhances the replication efficiency of SV40 DNA. (iii) Minor late promoters have been localized within the minimal replication origin and the 72-bp repeat. (iv) The minimal replication origin is not per se a constituent of the major late promoter; however, both the minimal replication origin and the 21-bp repeat are required for obtaining high levels of late gene expression observed at late times after infection by SV40. (v) The 72-bp repeat exerts a 4- to 5-fold enhancement of late promoter expression.
Monoclonal antibodies detect the conservation of muscarinic cholinergic receptor structure from Drosophila to human brain and detect possible structural homology with alpha 1-adrenergic receptors.
Authors: Venter J C , Eddy B , Hall L M , Fraser C M
Published: May 2006

Muscarinic cholinergic receptors isolated from Drosophila heads, rat and human brain, dog heart, and monkey ciliary muscle were examined for structural similarities/differences by utilizing isoelectric focusing, sodium dodecyl sulfate/polyacrylamide gel electrophoresis, and monoclonal antibody crossreactivity. Muscarinic receptors were affinity labeled with [3H]propylbenzilylcholine mustard and subjected to isoelectric focusing. Muscarinic receptors from each species focused with an isoelectric point of 5.9. The same proteins all migrated with an apparent molecular mass of 80,000 daltons on sodium dodecyl sulfate gels. Six hybridomas secreting monoclonal antibodies specific for muscarinic receptors were developed by using purified rat brain muscarinic receptors as the antigen. The six different monoclonal antibodies immunoprecipitated muscarinic receptors from all tissues and species tested, including human and Drosophila brains, with equal efficacy. These data indicate that muscarinic receptors are highly conserved over a considerable evolutionary period. One of the six muscarinic receptor monoclonal antibodies also immunoprecipitated rat liver alpha 1-adrenergic receptors. Furthermore, two out of five monoclonal antibodies raised against alpha 1-receptors immunoprecipitated muscarinic receptors. These data suggest that some degree of structural homology exists between muscarinic cholinergic receptors and alpha 1-adrenergic receptors.
Possible DNA-RNA tumor virus interaction in human lymphomas: expression of retroviral proteins in Ramos lymphoma lines is enhanced after conversion with Epstein-Barr virus.
Authors: Lasky R D , Troy F A
Published: May 2006

Epstein-Barr virus (EBV)-genome-negative human lymphoma lines, Ramos and BJAB, can be converted by EBV in vitro into EBV-genome-positive virus nonproducer lines. These cell lines have been used to identify surface antigens unique to EBV, with the expectation that such determinants might be related to the antigenic target responsible for EBV-specific immunosurveillance. Antisera prepared in rabbits immunized with either whole cells or purified plasma membranes were used in immunoblot experiments to analyze antigenic differences resulting from expression of the resident EBV genome. Unexpectedly, an increase in polypeptides of 32 and 70 kilodaltons was consistently observed in the EBV-converted Ramos lines. In contrast, these antigens were not expressed in BJAB or in its EBV-converted lines. These data suggested that p32 and gp70 might be murine leukemia virus (MuLV)-coded antigens because Ramos, but not BJAB, had been passaged in athymic nude mice during establishment of this cell line. This conclusion was confirmed by using antisera specific for MuLV p30 and gp70. Retroviral antigens were expressed constitutively at low levels in Ramos. Quantitative immunoblotting showed that EBV conversion of Ramos amplified the expression of MuLV proteins 3- to 5-fold. The molecular mechanism responsible for the enhanced expression is unknown. The biological relevance of this phenomenon is also not clear, but the interaction between a DNA and a RNA tumor virus in a Burkitt lymphoma line that carries both viruses may have important biological consequences in relation to retrovirus latency and tumor induction. These results also show that caution must be used when ascribing "uniqueness" to EBV-determined antigens, particularly in the Ramos lines. This warning extends also to the use of Ramos cell lines as immunogens, because immunization of rabbits elicited antibodies that recognized proteins of the same size as the retroviral antigens.
Steps in processing of the mitochondrial cytochrome oxidase subunit I pre-mRNA affected by a nuclear mutation in yeast.
Authors: Simon M , Faye G
Published: May 2006

In Saccharomyces cerevisiae, the mitochondrial gene encoding the subunit I of cytochrome c oxidase (oxi-3 gene) is interrupted by intervening sequences. In this report, a nuclear mutation [referred to as mss51 in Faye, G. & Simon, M. (1983) Cell 32, 77-87] that specifically affects the processing of oxi-3 pre-mRNA was further characterized. DNA probes covering each oxi-3 exon-intron boundary were individually hybridized to wild-type and mutant mitochondrial RNA. By a technique relying on the S1 nuclease resistance or sensitivity of the RNA X DNA hybrids thereof, we have shown which site needs the MSS51 gene product to be cleaved. The mutation in the MSS51 gene gave rise to a complex pattern of splicing: the third intron was excised efficiently but the first two introns remained bracketed by their flanking exons. Further, the fourth and fifth introns were only partially split from their common exon and remained fused to their upstream and downstream flanking exon, respectively. Several plausible roles for the MSS51 gene product are discussed.