bluuebunny/crossref_metadata_2025 huggingface dataset:
Choline kinase II is present only in nodules that synthesize stable peribacteroid membranes
Authors: Mellor Robert B. , Christensen Tove M. I. E. , Werner Dietrich
Published: May 2006

Host-cell cytoplasm from soybean plants infected with the peribacteroid membrane (PBM)-building
Rhizobium japonicum
strain 61-A-101 (effective, N
2
-fixing) had much higher choline kinase activity than cytoplasm from either uninfected tissue or tissue infected with the non-PBM-building (ineffective, non-N
2
-fixing) strain 61-A-24. Ion-exchange chromatography showed that both types of nodule and root tissue possessed constitutive choline kinase I activity that had a
K
m
for choline of ≈150 μM. The nodules of the effective symbiosis had another activity, choline kinase II (
K
m
= 81 μM). Nondenaturing and NaDodSO
4
electrophoresis revealed no multimeric subunit structure of the two enzyme forms but did show the molecular sizes for choline kinase I, 58-59 kDa, and choline kinase II, 60 kDa. Choline kinase I and II and pI values of 8.1 and 8.5, respectively, and two-dimensional gel electrophoresis of whole cytoplasm from control and infected tissue showed a spot corresponding to choline kinase II only in the case of the effective symbiosis, whereas both tissue types had spots corresponding to choline kinase I. Choline kinase II is presumed to be encoded by the plant as neither free-living nor symbiotic (bacteroid) forms of the prokaryote showed any choline kinase activity.
Multiple purine pathway enzyme activities are encoded at a single genetic locus in Drosophila.
Authors: Henikoff S , Keene M A , Sloan J S , Bleskan J , Hards R , Patterson D
Published: May 2006

The Drosophila melanogaster Gart locus, known from previous work to encode the enzyme activity phosphoribosylglycinamide formyltransferase (GART), specifies two alternatively processed mRNAs and two proteins. We introduced the entire Gart locus into a Drosophila tissue culture cell line in which the locus is active. The resulting cell clones contained numerous copies of the locus and overproduced both mRNAs and both expected proteins, thus markedly facilitating analysis of these molecules. We assayed extracts of the clones for the activities of 10 enzymes important for de novo purine synthesis and found that, in addition to GART, two other purine pathway activities, phosphoribosylamine-glycine ligase (phosphoribosylglycinamide synthetase, GARS) and phosphoribosylformylglycinamidine cyclo-ligase (phosphoribosylaminoimidazole synthetase, AIRS), are similarly overproduced. All three activities are present together on the larger overproduced protein. A smaller protein appears to possess only GARS activity. Therefore, alternative mRNA processing can allow cells to produce enzyme activities in forms that are either linked or unlinked to other activities.
A genomic clone encoding the alpha chain of the OKM1, LFA-1, and platelet glycoprotein IIb-IIIa molecules.
Authors: Cosgrove L J , Sandrin M S , Rajasekariah P , McKenzie I F
Published: May 2006

LFA-1, an antigen involved in cytolytic T lymphocyte-mediated killing, and Mac-1, the receptor for complement component C3bi, constitute a family of structurally and functionally related cell surface glycoproteins involved in cellular interactions. In both mouse and man, Mac-1 (OKM1) and LFA-1 share a common 95-kDa beta subunit but are distinguished by their alpha chains, which have different cellular distributions, apparent molecular masses (165 and 177 kDa, respectively), and peptide maps. We report the isolation of a genomic clone from a human genomic library that on transfection into mouse fibroblasts produced a molecule(s) reactive with monoclonal antibodies to OKM1, to LFA-1, and to platelet glycoprotein IIb-IIIa. This gene was cloned by several cycles of transfection of L cells with a human genomic library cloned in lambda phage Charon 4A and subsequent "rescue" of the lambda phage. Transfection with the purified recombinant lambda DNA yielded a transfectant that expressed the three human alpha chains of OKM1, LFA-1, and glycoprotein IIb-IIIa, presumably in association with the murine beta chain.
Cytogenetic "rogue" cells: what is their frequency, origin, and evolutionary significance?
Authors: Awa A A , Neel J V
Published: May 2006

Among 102,170 cultured lymphocytes obtained from 9818 individuals from Hiroshima, Japan, aged 9 to 37 years and scored for chromosomal abnormalities, 24 cells that exhibited an extreme degree of damage were encountered. The damage consists of multiple dicentric and even tricentric chromosomes, as well as numerous fragments, many with the appearance of "double minutes." The occurrence of these cells was not correlated with parental exposure to the atomic bomb, age, sex, year, or season. They were nonrandomly distributed by individual. Such cells were originally described in South American Indians and have also been recorded in inhabitants of the United States and the United Kingdom; this appears to be a world-wide phenomenon. Their cause remains unknown, and it is not known whether they occur in other somatic and germ-line cells. Should the latter be the case and should the least damaged of these cells occasionally successfully complete mitosis and meiosis, the possible role of such cells in oncogenesis and evolution must be considered.
Genetic instability in Drosophila melanogaster: the genetics of an MR element that makes complete P insertion mutations.
Authors: Green M M
Published: May 2006

An MR element that maps to a specific locus in chromosome 3 of Drosophila melanogaster has the unusual feature of producing complete P (a mobile element) sequence mutations at several X chromosome loci. This MR also increases the frequency of mitotic recombination. Evidence is given for the transposition of MR. The complete P insertion mutations are autonomously unstable and are capable of causing otherwise stable incomplete (defective) P insertion mutations to revert. These results complement the analysis of P element functions with a synthetic complete P derivative. The genetic basis for the mutational-mitotic recombinational components of "hybrid dysgenesis" is conveniently explicable in terms of MR elements present in the genomes of flies present in the wild.
Symbolic method in invariant theory
Authors: Rota Gian-Carlo , Stein Joel A.
Published: May 2006

A symbolic method based on an extension of the straightening algorithm is developed for the representation of joint invariants of symmetric and skew-symmetric tensors. For skew-symmetric tensors, the method holds over infinite fields of arbitrary characteristic.
Bacteriophage T4 DNA topoisomerase mediates illegitimate recombination in vitro.
Authors: Ikeda H
Published: May 2006

We have found that purified T4 DNA topoisomerase promotes recombination between two phage lambda DNA molecules in an in vitro system. In this cross, T4 DNA topoisomerase alone is able to catalyze the recombination and produce a linear monomer recombinant DNA that can be packaged in vitro. ATP is not required for this recombination, while oxolinic acid stimulates it. The recombinant DNA molecules contain duplications or deletions, and the crossovers take place between nonhomologous and nonspecific sequences of lambda DNA. Therefore, the recombination mediated by the T4 DNA topoisomerase is an illegitimate recombination that is similar to that mediated by Escherichia coli DNA gyrase. A model was proposed previously in which DNA gyrase molecules that are bound to DNA associate with each other and lead to the exchange of DNA strands through the exchange of DNA gyrase subunits. This model is also applicable to the recombination mediated by T4 DNA topoisomerase.
Origin of eukaryotic introns: a hypothesis, based on codon distribution statistics in genes, and its implications.
Authors: Senapathy P
Published: May 2006

A hypothesis for the origin of introns in eukaryotic genes is developed. By computer simulation it was found that the reading-frame lengths in a random nucleotide sequence are distributed in a negative exponential manner and that there exists an upper limit of about 200 codons in the length of the reading frames (RFs). These characteristics suggest that, if primordial DNA contained a random nucleotide sequence, the most primitive cells would have been under selective pressure to eliminate interfering stop codons in order to increase the length of RFs. Further, they indicate that the only possible way that a coding sequence that is considerably longer than 600 nucleotides could be derived from the short coding sequences occurring in a random sequence would be to splice the short coding sequences and to eliminate the stretches of sequences containing clusters of inframe stop codons. Thus, introns are suggested to be those stretches of sequences containing interfering stop codons that were originally earmarked in the first primitive cells to be eliminated in order to enable the coding for long polypeptides. Because the statistical characteristics of codon distributions in today's eukaryotic DNA sequences resemble closely those of a random sequence and because the upper limit in the length of RFs (200 codons) in a random sequence corresponds precisely to the observed maximum length of exons in today's eukaryotic genes (600 nucleotides), it is suggested that introns originated in the most primitive unicellular eukaryotes when they evolved from primordial sequences. The data from the prokaryotic gene sequences indicate that prokaryotic genes may have been derived originally from primitive unicellular eukaryotic genes by losing introns from them.
JC papovavirus large tumor (T)-antigen expression in brain tissue of acquired immune deficiency syndrome (AIDS) and non-AIDS patients with progressive multifocal leukoencephalopathy.
Authors: Stoner G L , Ryschkewitsch C F , Walker D L , Webster H D
Published: May 2006

Progressive multifocal leukoencephalopathy (PML) is a JC papovavirus infection of the central nervous system in immunocompromised patients. It is well established that demyelination in PML is caused by JC virus infection of oligodendroglia, but whether the nonstructural regulatory protein, large tumor (T) antigen, is detectable in infected human tissue was not known. Using a modification of the peroxidase-antiperoxidase technique, we found T antigen expressed in the nuclei of cells in virus-infected sites in five cases of PML studied, including two with acquired immune deficiency syndrome (AIDS). PML occurs in AIDS at a much higher frequency than in other immunosuppressive disorders, and PML in AIDS may represent a more severe form of JC virus infection of the central nervous system.
Metric rigidity theorems on Hermitian locally symmetric spaces
Authors: Mok Ngaiming
Published: May 2006

Let
X
= Ω/Γ be a compact quotient of an irreducible bounded symmetric domain Ω of rank ≥2 by a discrete group ω of automorphisms without fixed points. It is well known that the Kähler-Einstein metric
g
on
X
carries seminegative curvature (in the sense of Griffiths). I show that any Hermitian metric
h
on
X
carrying seminegative curvature must be a constant multiple of
g
. This can be applied to prove rigidity theorems of holomorphic maps from
X
into Hermitian manifolds (
Y, k
) carrying seminegative curvature. These results are also generalized to the case of quotients of finite volume. On the other hand, let (

X
c
, g
c

) be an irreducible compact Hermitian symmetric manifold of rank ≥2. Then

g
c

is Kähler and carries semipositive holomorphic bisectional curvature. I prove that any Kähler
h
on

X
c

carrying semipositive holomorphic bisectional curvature must be equal to

g
c

up to a constant multiple and up to a biholomorphic transformation of

X
c

.
Characterization of a cDNA coding for human factor VII.
Authors: Hagen F S , Gray C L , O'Hara P , Grant F J , Saari G C , Woodbury R G , Hart C E , Insley M , Kisiel W , Kurachi K
Published: May 2006

Factor VII is a precursor to a serine protease that is present in mammalian plasma. In its activated form, it participates in blood coagulation by activating factor X and/or factor IX in the presence of tissue factor and calcium. Clones coding for factor VII were obtained from two cDNA libraries prepared from poly(A) RNA from human liver and Hep G2 cells. The amino acid sequence deduced from the cDNAs indicates that factor VII is synthesized with a prepro-leader sequence of 60 or 38 amino acids. The mature protein that circulates in plasma is a single-chain polypeptide composed of 406 amino acids. The amino acid sequence analysis of the protein and the amino acid sequence deduced from the cDNAs indicate that factor VII is converted to factor VIIa by the cleavage of a single internal bond between arginine and isoleucine. This results in the formation of a light chain (152 amino acids) and a heavy chain (254 amino acids) that are held together by a disulfide bond. The light chain contains a gamma-carboxyglutamic acid (Gla) domain and two potential epidermal growth factor domains, while the heavy chain contains the serine protease portion of the molecule. Factor VII shows a high degree of amino acid sequence homology with the other vitamin K-dependent plasma proteins.
Resonance Raman evidence for oxygen exchange between the FeIV = O heme and bulk water during enzymic catalysis of horseradish peroxidase and its relation with the heme-linked ionization.
Authors: Hashimoto S , Tatsuno Y , Kitagawa T
Published: May 2006

Raman spectroscopic studies of compound II of horseradish peroxidase show that the oxygen atom in the FeIV = O group of the heme is rapidly exchanged in H2O at pH 7.0 but not in an alkaline solution (pH 11.0). This conclusion is based on studies of shift in the FeIV = O stretching mode of compound II in H2(18)O; further studies show that the FeIV = O heme is hydrogen-bonded to an amino acid residue of the protein in neutral solutions but not in the alkaline solution. Deprotonation of this residue takes place with the midpoint pH at 8.8 and accordingly corresponds to the so-called heme-linked ionization. It is concluded that this hydrogen-bonded proton plays an important part in the oxygen exchange mechanism. From this it seems clear that this hydrogen-bonded proton has an essential role in the acid/base catalysis of this enzyme and that alkaline deactivation of this enzyme can be attributed to the lack of a hydrogen-bonded proton at high pH.
Analysis of Agrobacterium tumefaciens virulence mutants in leaf discs.
Authors: Horsch R B , Klee H J , Stachel S , Winans S C , Nester E W , Rogers S G , Fraley R T
Published: May 2006

The leaf disc transformation system provides a simple means to score expression of various T-DNA markers within days of infection by Agrobacterium tumefaciens as well as long-term selection for growth of transformed callus and shoots. In this report, we describe the application of this system to evaluation of marker transfer and integration in a comprehensive set of defined avirulent mutants of A. tumefaciens. We conclude that virC is not essential when the T-DNA is present on a binary vector. The most likely explanation for this is that the virC gene product is involved in generation of a T-DNA intermediate.
Antagonism of phencyclidine action by metaphit in rat cerebellar Purkinje neurons: an electrophysiological study.
Authors: Wang Y , Palmer M , Freedman R , Hoffer B , Mattson M V , Lessor R A , Rice K C , Jacobson A E
Published: May 2006

Metaphit (1-[1-(3-isothiocyanatophenyl)-cyclohexyl]piperidine), a derivative of the psychotomimetic drug phencyclidine (PCP), is postulated to bind irreversibly to PCP receptors. We examined here the electrophysiological interactions of metaphit with PCP in rat cerebellar cortex, since a specific effect of PCP on cerebellar neuronal circuitry has been shown. Metaphit, applied locally to Purkinje neurons by micropressure ejection through multibarreled micropipettes, has a reversible depressant action lasting for 5-20 min. Following this, PCP-induced inhibition is blocked with no recovery despite repeated applications of PCP for over an hour. This blockade was not seen unless the dose of metaphit was sufficient to transiently depress Purkinje neuron discharge. Metaphit does not antagonize inhibitory effects of locally applied norepinephrine or gamma-aminobutyric acid. This electrophysiological data suggests that metaphit is an irreversible antagonist of PCP in the cerebellum.
Overlapping promoters and their control in Escherichia coli: the gal case.
Authors: Herbert M , Kolb A , Buc H
Published: May 2006

Two overlapping promoters compete for RNA polymerase in the region that controls the expression of the galactose operon in Escherichia coli. Kinetics of open complex formation at P1 and P2 can be followed through the rate of formation of two specific abortive transcripts. The corresponding forward kinetic constants appear to be identical over a wide range of enzyme concentrations and temperatures, indicating that the two processes are strongly coupled. We propose a scheme accounting for our observations. In a first step, the competition between the two sites is a simple kinetic process, involving the "on" rate constants. In a second step, a slow reequilibration occurs, implicating the "off" rate constants and the conversion of one open complex to the other through a set of closed complexes. The first step is clearly affected when the complex between cyclic AMP and its receptor is bound at the activator site. An estimate of the various rate constants describing open complex formation at P1 and P2 is provided, as well as a qualitative description of the effect of the activator complex on these two pathways.
Mixtec social “credit rating”—The particular versus the universal in one emic world view
Authors: Pike Kenneth L.
Published: May 2006

That a society controls, to a greater or lesser extent, the behavior of its members is a universal; but the methods, the particulars of that control, vary from one culture to another. It is also, I would guess, a universal that in all societies people value respectability granted to them. If this is lost, the losers may become ineligible for many of the responsibilities and privileges of that society or subsociety. In this article, three ways are discussed by which such “credit rating” can be lost—as told by one member of a particular geographical subsection of the Mixtec society, as he has seen and lived it. Difficulties arose for some of its members, for example, from their refusal to vote with the majority, or from failure to care for children adequately in terms of grandparents' evaluation of the parents' responsibility, or from moral delinquency as judged by other members. Such internal cultural standards may be labeled “emic.”
Cloning and cDNA sequence of the regulatory subunit of cAMP-dependent protein kinase from Dictyostelium discoideum.
Authors: Mutzel R , Lacombe M L , Simon M N , de Gunzburg J , Veron M
Published: May 2006

cDNA clones encoding the regulatory subunit of the cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) from Dictyostelium discoideum were isolated by immunoscreening of a cDNA library constructed in the expression vector lambda gt11. High-affinity cAMP-binding activity was detected in extracts from bacteria lysogenized with these clones. Nucleotide sequence analysis of three overlapping clones allowed the determination of a 1195-base-pair cDNA sequence coding for the entire regulatory subunit and containing nontranslated 5' and 3' sequences. The open reading frame codes for a protein of 327 amino acids, with molecular weight 36,794. The regulatory subunit from Dictyostelium shares a high degree of homology with its mammalian counterparts, but is lacking the NH2-terminal domain required for the association of regulatory subunits into dimers in other eukaryotes. On the basis of the comparison of the regulatory subunits from Dictyostelium, yeast, and bovine tissues, a model for the evolution of these proteins is proposed.
Biosynthesis, glycosylation, and partial N-terminal amino acid sequence of the T-cell-activating protein TAP.
Authors: Reiser H , Coligan J , Benacerraf B , Rock K L
Published: May 2006

We have characterized the TAP molecule, an Ly-6 linked T-cell-activating glycoprotein. The three TAP bands that are precipitated from metabolically labeled cells display a common migration pattern in isoelectric focusing/NaDodSO4/PAGE gels and have common N-terminal sequences. This sequence is rich in cysteine and is homologous to that previously reported for the Ly-6.1E antigen. We, therefore, compared TAP and Ly-6.1E biochemically and found them to be structurally distinct. Given the role of TAP in T-cell activation, we further studied whether the molecule was phosphorylated. We have not found evidence for phosphorylation of the TAP protein. The carbohydrates present on the TAP molecule are resistant to peptide N-glycosidase F in vitro and tunicamycin in vivo. The upper band of the TAP triplet is susceptible to treatment with trifluoromethanesulfonic acid and thus seems to be of the O-linked rather than of the N-linked variety. The biosynthetic processing of TAP was studied in pulse-chase experiments. The middle band of the TAP triplet appears to be the earliest detectable species. Its conversion to the O-linked high molecular weight species can be blocked by monensin.
Lymphocyte major histocompatibility complex-encoded class II structures may act as sperm receptors.
Authors: Ashida E R , Scofield V L
Published: May 2006

Human sperm and blood cells were cocultured in vitro to determine whether specific interactions occur between gametes and blood cells. Evidence for cell type-specific sperm binding and penetration of lymphocytes is presented together with findings that suggest that either or both events involve major histocompatibility complex-encoded class II molecules on lymphocytes and a sperm ligand that is immunoreactive with antibodies to T-cell surface antigen T4. Involvement of HLA-DR is suggested by the pattern of sperm interactions with HLA-DR-positive and -negative cells and by inhibition of sperm binding to HLA-DR-positive cells by a monoclonal antibody that identifies a nonpolymorphic determinant on the HLA-DR molecule. That the complementary sperm ligand may be a T4-like structure is suggested by specific inhibition of sperm-lymphocyte binding with monoclonal antibodies OKT4 and OKT4A. The results are discussed in terms of possible roles for immunoglobulin-related structures in human fertilization and in the sexual transmission of the acquired immunodeficiency syndrome.
Crystal structure of hexagonal MnAl
4
Authors: Pauling Linus
Published: May 2006

A structure is proposed for the hexagonal form of MnAl
4
, with
a
H
= 28.4 Å and
c
H
= 12.43 Å, on the basis of a high-resolution electron micrograph and comparison with crystals of known structures. The proposed structure involves seven 104-atom complexes of 20 Friauf polyhedra, sharing some atoms with one another. It is closely related to the 23.36-Å cubic structure of MnAl
4
and to the 14.19-Å cubic structure of Mg
32
(Al,Zn)
49
.
Transcriptional (p40x) and post-transcriptional (p27x-III) regulators are required for the expression and replication of human T-cell leukemia virus type I genes.
Authors: Inoue J , Yoshida M , Seiki M
Published: May 2006

The pX sequence of human T-cell leukemia virus type I codes for three products: p40x, p27x-III, and p21x-III. p40x is a transcriptional trans-activator that activates not only the viral long terminal repeat but also cellular genes for interleukin 2 and its receptor. p27x-III and p21x-III are not required for transcriptional activation, and their functions were unknown. Cotransfection experiments with defective human T-cell leukemia virus type I proviruses and various pX expression plasmids revealed that p27x-III, in addition to p40x, was required for gag gene expression. Furthermore, it was shown that p27x-III induced accumulation of a high level of unspliced viral gag mRNA. These results indicate that p27x-III is a post-transcriptional modulator of viral RNA whose transcription has been fully activated by p40x.
Selective resistance of bone marrow-derived hemopoietic progenitor cells to gliotoxin.
Authors: Müllbacher A , Hume D , Braithwaite A W , Waring P , Eichner R D
Published: May 2006

The fungal metabolite gliotoxin at low concentrations prevents mitogen stimulation of mature lymphocytes as a result of gliotoxin-induced genomic DNA degradation. Bone marrow, on the other hand, contains a subpopulation of cells resistant to gliotoxin at similar concentrations. This population includes the hemopoietic progenitor cells that grow in vitro in response to appropriate colony-stimulating factors and cells that form colonies in the spleens of lethally irradiated recipients. Gliotoxin treatment of lymph node cell-enriched bone marrow significantly delayed the onset of graft-versus-host disease in fully allogeneic bone marrow chimeras.
Transmission in NFS/N mice of the heritable spongiform encephalopathy associated with the gray tremor mutation.
Authors: Hoffman P M , Rohwer R G , MacAuley C , Bilello J A , Hartley J W , Morse H C
Published: May 2006

It has been shown that the autosomal recessive mutation, gray tremor (gt) was associated in the homozygous state (gt/gt) with a rapidly fatal spongiform encephalopathy. Heterozygotes (+/gt) developed mild asymptomatic spongiform brain lesions as did recipient inbred mice inoculated with gt/gt brain homogenates, some of whom also showed behavioral abnormalities [Sidman, R. L., Kinney, H. C. & Sweet, H. O. (1985) Proc. Natl. Acad. Sci. USA 82, 253-257]. In these studies, inbred NFS/N mice inoculated intracerebrally at birth or as adults with gt/gt or first passage gt brain homogenates developed a progressive disease characterized by tremor, ataxia, and spasticity. The symptoms were milder and more slowly progressive than in the gt/gt homozygote, in the paralytic syndrome that followed neonatal inoculation of NFS/N mice with a wild murine leukemia virus (Cas-Br-M MuLV), or in the rapidly progressive ataxia and terminal bradykinesia that followed scrapie inoculation of NFS/N mice. The noninflammatory spongiform encephalopathy in affected NFS/N mice resembled that observed in gt/gt homozygotes, +/gt heterozygotes, and asymptomatic recipient inbred mice inoculated with gt/gt brain homogenates. Neither infectious MuLV nor MuLV proteins were detected in gt/gt brain homogenates or in affected recipient mouse brains. Scrapie-associated fibrils, readily identifiable in subcellular fractions of brains from scrapie-inoculated NFS/N mice, were not detected in similar brain fractions from NFS/N mice inoculated with gt brain homogenates. These results confirm and extend the suggestion that gt spongiform encephalopathy has both heritable and transmissible properties. Moreover, the transmissible agent of gt disease differs from both Cas-Br-M MuLV and scrapie in its disease-inducing properties in NFS/N mice. The capacity of NFS/N mice to express transmitted gt encephalopathy as clinical disease, to rapidly express Cas-Br-M MuLV spongiform encephalomyelopathy, and to develop mouse-adapted scrapie after a very short incubation time suggest a distinct sensitivity of NFS/N mice to transmissible spongiform encephalopathy.
The narL gene product activates the nitrate reductase operon and represses the fumarate reductase and trimethylamine N-oxide reductase operons in Escherichia coli.
Authors: Iuchi S , Lin E C
Published: May 2006

Escherichia coli, which can utilize O2, nitrate, fumarate, or trimethylamine N-oxide (Me3NO) as terminal electron acceptor, preferentially utilizes the one with the highest redox potential. Thus O2 prevents induction of nitrate, fumarate, and Me3NO reductases, and nitrate curtails the induction of fumarate and Me3NO reductases. Under anaerobic conditions the narL gene product, in the presence of nitrate, is known to activate transcription of the narC operon, which encodes nitrate reductase. This study shows that the same product plays a role in the repression by nitrate of the operons (frd and tor) that encode fumarate and Me3NO reductases. In contrast, the anaerobic repression of ethanol dehydrogenase by nitrate does not require the narL product. Expression of narL does not require the fnr gene product, a pleiotropic activator that is required for full expression of narC, frd, and tor.
Alternative perspective on photosynthetic yield and enhancement
Authors: Warner J. W. , Berry R. Stephen
Published: May 2006

In the traditional Z scheme of photosynthesis the Emerson effects of red drop (decline in yield of photosynthesis in far-red light) and enhancement (of far-red yield by supplementary short-wavelength light) are taken to be evidence for the coupling in series of two photosystems that absorb unsymmetrically in the far-red region of the spectrum. An alternative explanation for red drop and enhancement is proposed here that does not invoke the series-coupling hypothesis. It is suggested that the Emerson effects may be due to the drop in intensity of radiation from sample absorption, which causes a photochemical loss when the reaction shuts off at depth in the medium. The effects of oxygen, carbon dioxide, and temperature on the yield may also be interpreted in terms of this model.
Increased radiation-induced transformation in C3H/10T1/2 cells after transfer of an exogenous c-myc gene.
Authors: Sorrentino V , Drozdoff V , Zeitz L , Fleissner E
Published: May 2006

C3H/10T1/2 cells were infected with a retroviral vector expressing a mouse c-myc oncogene and a drug-selection marker. The resulting cells, morphologically indistinguishable from C3H/10T1/2, displayed a greatly enhanced sensitivity to neoplastic transformation by ionizing radiation or by a chemical carcinogen. Constitutive expression of myc therefore appears to synergize with an initial carcinogenic event, providing a function analogous to a subsequent event that apparently is required for the neoplastic transformation of these cells. This cell system should prove useful in exploring early stages in radiation-induced transformation.
Field tests of interspecific resource-based competition among phytoplankton
Authors: Carney Heath J.
Published: May 2006

The hypothesis that interspecific resource-based competition caused the spring and summer vertical segregation of phytoplankton species was tested in Lake Tahoe (California/Nevada). Two species (
Cyclotella glomerata
Bachmann and
Synedra radians
Kütz.) became dominant at different depth intervals (0-30 m and 60-90 m, respectively). Experimental transplants of assemblages between depths demonstrated asymmetrical competition. In the phosphate-limited region near the surface, growth of
S. radians
declined in the presence of
C. glomerata
. However, growth of
C. glomerata
was not affected by the presence of
S. radians
in the light-limited region at depth. This study provides field verification of resource-based competition theory.
Microcell-mediated transfer of a single human chromosome complements xeroderma pigmentosum group A fibroblasts.
Authors: Schultz R A , Saxon P J , Glover T W , Friedberg E C
Published: May 2006

Chromosomes from an immortalized aneuploid human fibroblast cell line were randomly tagged with the selectable marker neo by transfection with the plasmid pSV2neo. Somatic cell fusions between transfected human cells and mouse A9 cells generated pools of G418-resistant human-mouse hybrid clones containing various numbers of human chromosomes. Microcell-mediated chromosome transfer from the hybrid pools to xeroderma pigmentosum complementation group A (XP-A) cells in culture and selection for G418-resistant colonies resulted in the identification of XP cells with enhanced resistance to ultraviolet radiation. Screening of subclones from selected pools of human-mouse hybrids facilitated the identification of hybrids containing a single neo-tagged human chromosome. Transfer of this chromosome to XP-A cells (but not to XP-F or XP-C cells) results in enhanced resistance to ultraviolet light and enhanced excision repair capacity. The identification of a single human chromosome that complements the phenotype of XP-A cells in culture provides the potential for genetic mapping of the complementing gene and for its isolation by molecular cloning.
N-methyl-D-aspartate receptor antagonist desegregates eye-specific stripes.
Authors: Cline H T , Debski E A , Constantine-Paton M
Published: May 2006

The optic tecta of surgically produced three-eyed tadpoles were chronically exposed to the N-methyl-D-aspartate (NMDA) receptor antagonist aminophosphonovaleric acid (APV), or to NMDA itself, to assess the influence of NMDA receptor/channels on the eye-specific segregation of retinal ganglion cell (RGC) terminals that occurs whenever two retinas innervate one tectal lobe. Exposure of the tectum to the active isomer of APV produces desegregation of the RGC terminals without blocking electrical activity in the afferents or altering their terminal arbor morphology. Exposure to the inactive isomer of APV causes no perturbation of the normal stripe pattern. APV-induced desegregation is completely reversible within 2 weeks of removal of the APV. In addition, exposure of the optic tectum to NMDA results in stripes with sharper borders and fewer forks and fusions than untreated animals. These results suggest that the NMDA receptor/channel plays a role in eye-specific segregation in the three-eyed tadpole.
Molecular cloning and sequence of a cDNA coding for bovine lipoprotein lipase.
Authors: Senda M , Oka K , Brown W V , Qasba P K , Furuichi Y
Published: May 2006

Lipoprotein lipase (LPL; triacylglycero-protein acylhydrolase, EC 3.1.1.34) was purified from bovine milk. Synthetic oligonucleotides were prepared, based on the amino acid sequences of three peptides obtained from partial digestion of purified LPL, and were used as probes to isolate cDNA clones for LPL mRNA from a bovine mammary gland. One of the clones, pLPL-49R2, contains an insert cDNA (49R2) of about 3.2 kilobases (kb) that hybridizes to all three probes and encodes a polypeptide that includes the NH2-terminal sequence of bovine LPL reported recently [Ben-Avram, C. M., Ben-Zeev, O., Lee, T. D., Hagga, K., Shively, J. E., Goers, J., Pedersen, M. E., Reeve, J. R. & Schotz, M. C. (1986) Proc. Natl. Acad. Sci. USA 83, 4185-4189]. Complete nucleotide sequence analysis revealed that cDNA insert 49R2 contains the entire coding region for LPL as well as a 3' untranslated region of about 1.6 kb. The predicted amino acid sequence indicates that bovine LPL is a hydrophilic protein consisting of 450 amino acids (Mr 50,548) in its unglycosylated form. Blot hybridization analysis of poly(A)+ mRNA from bovine mammary gland demonstrated that there are at least three sizes of LPL mRNAs--3.2, 2.5, and 1.7 kb--with the 2.5-kb mRNA being the most abundant. Restriction endonuclease mapping of other cDNA clones suggested that the variation in mRNA size results from differential utilization of polyadenylylation signals during mRNA processing.
DNase I-hypersensitive sites associated with expression and hormonal regulation of mouse C4 and Slp genes.
Authors: Hemenway C , Robins D M
Published: May 2006

There are four major regions of DNase I hypersensitivity in the 5' regions of the genes for the murine fourth component of complement (C4) and its homologous neighbor, Slp (sex-limited protein). Hypersensitivity around the start site of transcription and approximately equal to 0.5 kilobases upstream correlates qualitatively with expression of these genes. Two hypersensitive sites, at -2.3 and -2.0 kilobases, map specifically to the Slp gene and correlate with its hormonal regulation. That is, these sites are more prominent in male liver chromatin and become more apparent in chromatin from females treated with testosterone. Further, these sites are established in males to a greater extent than in females prior to expression of Slp and may reflect gene-commitment events. Comparison of chromatin from mouse strains differing in C4 and Slp alleles indicates that the four regions of hypersensitivity may be necessary but are not sufficient for high levels of expression.
Expression of members of immunoglobulin gene family in somatic cell hybrids between human B and T cells.
Authors: Kozbor D , Burioni R , Ar-Rushdi A , Zmijewski C , Croce C M
Published: May 2006

Somatic cell hybrids were obtained between human T and B cells and tested for the expression of differentiated traits of both cell lineages. The T-cell parent SUP-T1 is CD3-, CD4+, CD1+, CD8+, is weakly positive for HLA class I determinants, and has an inversion of chromosome 14 due to a site-specific recombination event between an immunoglobulin heavy-chain variable gene and the joining segment of the T-cell receptor alpha chain. The B-cell parent, the 6-thioguanine- and ouabain-resistant mutant GM1500, is a lymphoblastoid cell line that secretes IgG2, kappa chains, and expresses B1, B532, and HLA class I and II antigens. All hybrids expressed characteristics of B cells (Ig+, B1+, B532+, EBNA+, HLA antigens), whereas only CD4 among the T-cell markers was expressed. The level of T-cell receptor beta-chain transcript was greatly reduced and no RNA of the chimeric T-cell receptor alpha-chain joining segment-immunoglobulin heavy-chain variable region was detected. Southern blot analysis indicated that absence of T-cell differentiation markers in the hybrids was not due to chromosomal loss. Rather, some B-cell-specific factor present in the hybrids may account for the suppression.
Site-directed mutagenesis of an HLA-A3 gene identifies amino acid 152 as crucial for major-histocompatibility-complex-restricted and alloreactive cytotoxic-T-lymphocyte recognition.
Authors: Cowan E P , Jelachich M L , Coligan J E , Biddison W E
Published: May 2006

Major histocompatibility complex-restricted and alloreactive cytotoxic T lymphocytes (CTL) can discriminate between the HLA-A3.1 and HLA-A3.2 antigens. HLA-A3.1 and the rare variant HLA-A3.2 have been shown to differ by two amino acids in the alpha 2 domain at positions 152 (A3.1, glutamic acid; A3.2, valine) and 156 (A3.1, leucine; A3.2, glutamine). To determine the structural basis for the ability of CTL to differentiate A3.1 from A3.2, two site-directed mutants of the HLA-A3.2 gene were produced, 152A3.1-156A3.2 and 152A3.2-156A3.1, that have the indicated codons for positions 152 and 156. These mutated HLA-A3 genes, as well as the nonmutated HLA-A3.1 and HLA-A3.2 genes, were then transfected into the murine cell line P815-HTR and used as targets for human CTL. Influenza virus-specific HLA-A3.1-restricted CTL lysed virus-infected P815 cells transformed with the HLA-A3.1 and 152A3.1-156A3.2 genes, but not P815 cells transformed with the HLA-A3.2 and 152A3.2-156A3.1 genes. HLA-A3.2-allospecific CTL lysed the P815 cells transformed with the HLA-A3.2 and 152A3.2-156A3.1 genes but did not lyse P815 cells transformed with the HLA-A3.1 or 152A3.1-156A3.2 genes. Thus, a single amino acid change at position 152, substituting valine for glutamic acid and thereby introducing a charge difference, produces major structural changes in the epitopes recognized by major histocompatibility complex-restricted and alloreactive CTL.
Down-regulation of rat kidney calcitonin receptors by salmon calcitonin infusion evidenced by autoradiography.
Authors: Bouizar Z , Rostène W H , Milhaud G
Published: May 2006

In treating age-related osteoporosis and Paget disease of bone, it is of major importance to avoid an escape phenomenon that would reduce effectiveness of the treatment. The factors involved in the loss of therapeutic efficacy with administration of large pharmacological doses of the hormone require special consideration. Down-regulation of the hormone receptors could account for the escape phenomenon. Specific binding sites for salmon calcitonin (sCT) were characterized and localized by autoradiography on rat kidney sections incubated with 125I-labeled sCT. Autoradiograms demonstrated a heterogenous distribution of 125I-labeled sCT binding sites in the kidney, with high densities in both the superficial layer of the cortex and the outer medulla. Infusion of different doses of unlabeled sCT by means of Alzet minipumps for 7 days produced rapid changes in plasma calcium, phosphate, and magnesium levels, which were no longer observed after 2 or 6 days of treatment. Besides, infusion of high doses of sCT induced down-regulation of renal sCT binding sites located mainly in the medulla, where calcitonin (CT) has been shown to exert its physiological effects on water and ion reabsorption. These data suggest that the resistance to high doses of sCT often observed during long-term treatment of patients may be the consequence of not only bone-cell desensitization but also down-regulation of CT-sensitive kidney receptor sites.
Oligo(alpha-deoxynucleotide)s covalently linked to intercalating agents: differential binding to ribo- and deoxyribopolynucleotides and stability towards nuclease digestion.
Authors: Thuong N T , Asseline U , Roig V , Takasugi M , Hélène C
Published: May 2006

An octathymidylate was synthesized with the alpha anomer of thymidine instead of the naturally occurring beta anomer. This oligonucleotide binds to complementary sequences containing beta-nucleosides. Binding to ribose-containing oligomers and polymers is much stronger than binding to deoxyribose-containing analogs. A derivative of acridine (9-amino-6-chloro-2-methoxyacridine) was covalently attached either to the 5' phosphate or to the 3' phosphate of the alpha-octathymidylate. A pentamethylene linker was used to bridge the phosphate group and the 9-amino group of the acridine derivative. In both cases the complexes with the complementary sequences were strongly stabilized due to the additional binding energy provided by intercalation of the acridine ring within the miniduplex structure formed by the oligonucleotide with its target sequence. The acridine-substituted alpha-oligothymidylates did not lose their discrimination between ribose and deoxyribose-containing complementary sequences. The alpha-oligothymidylates were much more resistant towards endonucleases than their beta analogs, independently of whether they were linked to the acridine derivative. Acridine substitution provided additional protection against the corresponding exonucleases. alpha-Oligodeoxynucleotides covalently linked or not to intercalating agents represent families of molecules that open possibilities to block mRNA translation or viral RNA expression in vitro and in vivo.
Molecular genetic approach to human meningioma: loss of genes on chromosome 22.
Authors: Seizinger B R , de la Monte S , Atkins L , Gusella J F , Martuza R L
Published: May 2006

A molecular genetic approach employing polymorphic DNA markers has been used to investigate the role of chromosomal aberrations in meningioma, one of the most common tumors of the human nervous system. Comparison of the alleles detected by DNA markers in tumor DNA versus DNA from normal tissue revealed chromosomal alterations present in primary surgical specimens. In agreement with cytogenetic studies of cultured meningiomas, the most frequent alteration detected was loss of heterozygosity on chromosome 22. Forty of 51 patients were constitutionally heterozygous for at least one chromosome 22 DNA marker. Seventeen of the 40 constitutionally heterozygotic patients (43%) displayed hemizygosity for the corresponding marker in their meningioma tumor tissues. Loss of heterozygosity was also detected at a significantly lower frequency for markers on several other autosomes. In view of the striking association between acoustic neuroma and meningioma in bilateral acoustic neurofibromatosis and the discovery that acoustic neuromas display specific loss of genes on chromosome 22, we propose that a common mechanism involving chromosome 22 is operative in the development of both tumor types. Fine-structure mapping to reveal partial deletions in meningiomas may provide the means to clone and characterize a gene (or genes) of importance for tumorigenesis in this and possibly other clinically associated tumors of the human nervous system.
Simultaneous measurement of hormone release and secretagogue binding by individual pituitary cells.
Authors: Smith P F , Neill J D
Published: May 2006

The quantitative relationship between receptor binding and hormone secretion at the single-cell level was investigated in the present study by combining a reverse hemolytic plaque assay for measurement of luteinizing hormone (LH) secretion from individual pituitary cells with an autoradiographic assay of 125I-labeled gonadotropin-releasing hormone (GnRH) agonist binding to the same cells. In the plaque assay, LH secretion induces complement-mediated lysis of the LH-antibody-coated erythrocytes around the gonadotropes, resulting in areas of lysis (plaques). LH release from individual gonadotropes was quantified by comparing radioimmunoassayable LH release to hemolytic area in similarly treated cohort groups of cells; plaque area was linearly related to the amount of LH secreted. Receptor autoradiography was performed using 125I-labeled GnRH-A (a superagonist analog of GnRH) both as the ligand and as the stimulant for LH release in the plaque assay; the developed silver grains appearing over cells in the center of plaques were measured microscopically. The grains appeared to represent specific and high-affinity receptors for GnRH because no pituitary cells other than gonadotropes bound the labeled ligand and grain development was progressively inhibited by coincubation with increasing doses of unlabeled GnRH-A. Despite high correlations between mean grain number and mean plaque area in dose-response curves, the correlation coefficients for these parameters were low (range 0.02-0.38) in the individual cells comprising these groups. We conclude that GnRH receptor number for any individual gonadotrope is a weak determinant of the amount of LH it can secrete; nevertheless, full occupancy of all its GnRH receptors is required for any gonadotrope to reach its full LH-secretory capacity. Apparently the levels of other factors comprising the steps along the secretory pathway determine the secretory capacity of an individual cell.
Escherichia coli contains a soluble ATP-dependent protease (Ti) distinct from protease La.
Authors: Hwang B J , Park W J , Chung C H , Goldberg A L
Published: May 2006

The energy requirement for protein breakdown in Escherichia coli has generally been attributed to the ATP-dependence of protease La, the lon gene product. We have partially purified another ATP-dependent protease from lon-cells that lack protease La (as shown by immunoblotting). This enzyme hydrolyzes [3H]methyl-casein to acid-soluble products in the presence of ATP and Mg2+. ATP hydrolysis appears necessary for proteolytic activity. Since this enzyme is inhibited by diisopropyl fluorophosphate, it appears to be a serine protease, but it also contains essential thiol residues. We propose to name this enzyme protease Ti. It differs from protease La in nucleotide specificity, inhibitor sensitivity, and subunit composition. On gel filtration, protease Ti has an apparent molecular weight of 370,000. It can be fractionated by phosphocellulose chromatography or by DEAE chromatography into two components with apparent molecular weights of 260,000 and 140,000. When separated, they do not show proteolytic activity. One of these components, by itself, has ATPase activity and is labile in the absence of ATP. The other contains the diisopropyl fluorophosphate-sensitive proteolytic site. These results and the similar findings of Katayama-Fujimura et al. [Katayama-Fujimura, Y., Gottesman, S. & Maurizi, M. R. (1987) J. Biol. Chem. 262, 4477-4485] indicate that E. coli contains two ATP-hydrolyzing proteases, which differ in many biochemical features and probably in their physiological roles.