bluuebunny/crossref_metadata_2025 huggingface dataset:
Goodpasture antigen of the glomerular basement membrane: localization to noncollagenous regions of type IV collagen.
Authors: Wieslander J , Barr J F , Butkowski R J , Edwards S J , Bygren P , Heinegård D , Hudson B G
Published: May 2006

The glomerular basement membrane antigen in Goodpasture syndrome is a collagenase-resistant molecule with a monomer molecular weight of about 26,000. Type IV collagen isolated from glomerular basement membrane contains collagenase-resistant sequences within its structure. Polyacrylamide gel electrophoresis, enzyme-linked immunosorbent assay, and chemical analysis were used to demonstrate that the collagenase-resistant sequences of type IV collagen contain Goodpasture antigen.
Purification and NH2-terminal amino acid sequencing of the beta subunit of a human T-cell antigen receptor.
Authors: Acuto O , Fabbi M , Smart J , Poole C B , Protentis J , Royer H D , Schlossman S F , Reinherz E L
Published: May 2006

To obtain information about the structural basis for T-cell antigen recognition, a T3-associated Ti receptor molecule was isolated from crude membranes of the REX human thymic tumor line and purified by affinity chromatography with an anti- clonotypic monoclonal antibody in conjunction with preparative gel electrophoresis. NH2-terminal amino acid sequencing of the beta subunit unambiguously identified the amino acids in positions 2-12. Comparative protein sequence analysis by computer search demonstrated that this Ti beta sequence bore weak, but definite, homology to the first framework of the variable region of human lambda light chain. Anti-sera to a synthetic peptide corresponding to positions 2-11 precipitated the denatured Ti beta subunit from REX, thus confirming the above sequence. This information suggests that the Ti beta subunit is distantly related to human immunoglobulin lambda light chain and, moreover, should be of use in the molecular cloning of the Ti beta gene.
Human T-cell leukemia virus-associated membrane antigens: identity of the major antigens recognized after virus infection.
Authors: Lee T H , Coligan J E , Homma T , McLane M F , Tachibana N , Essex M
Published: May 2006

Specific antibodies to cell membrane antigens found on human T-cell leukemia virus (HTLV)-infected cells have been detected in Japanese patients with adult T-cell leukemia/lymphoma and in asymptomatic carriers, using a live cell-membrane immunofluorescence assay. Reactivity of the positive antisera was analyzed using radioimmunoprecipitation and NaDodSO4/PAGE with the HTLV-infected tumor cell line Hut 102 (clone B2). The major cell-associated antigens identified include two glycoproteins of approximately equal to 61 and 45 kDa, which appear to be the most immunogenic species in exposed people, a nonglycosylated species of 42 kDa, and four additional species that contain gag gene-encoded antigens with sizes ranging from 19 to 55 kDa. The two glycoproteins ( gp61 and gp45 ) are encoded, at least in part, by the env gene of HTLV as evidenced by amino acid sequence analysis.
Identification of the second gonadotropin-releasing hormone in chicken hypothalamus: evidence that gonadotropin secretion is probably controlled by two distinct gonadotropin-releasing hormones in avian species.
Authors: Miyamoto K , Hasegawa Y , Nomura M , Igarashi M , Kangawa K , Matsuo H
Published: May 2006

A new peptide having gonadotropin-releasing activity distinct from the known luteinizing hormone-releasing hormones ( LHRHs ) has been identified in a chicken hypothalamic extract. The existence of [Gln8]LHRH in avian hypothalamus has been reported previously. The new molecular species of gonadotropin-releasing activity, named chicken gonadotropin-releasing hormone II (chicken GnRH-II), has been isolated recently in a yield of 7 micrograms, starting from 10,000 chicken hypothalami. Structural analyses have been performed on the peptide fragments derived from chymotryptic and thermolytic digests of chicken GnRH-II by amino acid analyses and terminal analyses. The full sequence of chicken GnRH-II has been determined to be: pGlu-His-Trp-Ser-His-Gly-Trp-Tyr-Pro-Gly-NH2. A synthetic decapeptide with the above sequence was verified to be chromatographically identical to natural chicken GnRH-II. For further structural confirmation, chymotryptic and thermolytic peptides from synthetic and natural chicken GnRH-II also were identified. Thus, the structure of chicken GnRH-II has been definitely established. The gonadotropin-releasing potency of chicken GnRH-II was about 32% of that of mammalian LHRH and 8 times more potent than chicken LHRH, as estimated from the bioassay with rat anterior pituitary cells. Our results indicate that gonadotropin secretion is probably controlled by two distinct GnRHs , at least in avian species.
Model of specific complex between catabolite gene activator protein and B-DNA suggested by electrostatic complementarity.
Authors: Weber I T , Steitz T A
Published: May 2006

Calculation of the electrostatic potential energy surfaces of Escherichia coli catabolite gene activator protein (CAP) dimer suggests a model for the complex between CAP and a specific DNA sequence. The positive electrostatic charge density of CAP lies on the two COOH-terminal domains and about 20-30 A from the molecular 2-fold axis. Assuming that the 2-fold axes of the CAP dimer and the DNA to which it binds are coincident, the positions of the positive electrostatic potential surfaces strongly suggest the rotational orientation of the DNA relative to the protein. A specific complex between CAP and its DNA binding site in the lac operon has been built with the DNA in this orientation. The amino ends of the two protruding F alpha-helices interact in successive major grooves of the DNA. Four side chains emanating from each F helix can form hydrogen bonds with the exposed edges of four bases in the major groove. Electrostatic considerations as well as the necessity to make interactions between CAP and a DNA site as much as 20 base pairs long require us to bend or kink the DNA. In our model of CAP complexed with B-DNA, as with those proposed for Cro and lambda cI repressors, the protruding second helices of the two-helix motif from both subunits interact in successive major grooves of B-DNA. However, unlike Cro and similar to lambda cI, the protruding alpha-helices are nearly parallel to the bases rather than the groove.
Induction of gamma-interferon activity by elevated temperatures in human B-lymphoblastoid cell lines.
Authors: Taylor M W , Long T , Martinez-Valdez H , Downing J , Zeige G
Published: May 2006

When the human B-lymphoblastoid cell lines GM3299 and GM4408 were heat shocked, elevated levels of antiviral activity were detected. This antiviral activity has been characterized as the result of de novo synthesis of interferon (IFN) gamma. Levels of antiviral activity as high as 20,000 units/ml per 10(6) cells were found after heat treatment. By using anti-IFN-alpha and anti-IFN-gamma sera, it was shown that the high antiviral activity in GM3299 after heat shock is due to a synergistic effect of constitutively produced IFN-alpha and induced IFN-gamma.
Multiple regulatory signals in the control region of the Escherichia coli carAB operon.
Authors: Bouvier J , Patte J C , Stragier P
Published: May 2006

The first reaction in pyrimidine and arginine biosynthesis in Escherichia coli is catalyzed by a single enzyme, carbamoyl-phosphate synthetase (EC 6.3.5.5), the product of the carAB operon. Expression of this operon is cumulatively repressed by arginine and pyrimidines. The nucleotide sequence of the carAB control region was determined and transcriptional starts were localized. Two adjacent promoters, 70 base pairs apart, appear to be used in vivo, the downstream one overlapping a typical arginine operator. The absence of any attenuation-like sequence excludes such a mechanism for pyrimidine-mediated repression. Various fragments of the carA promoter-proximal region were fused in vitro with the lacZ gene. Results obtained with these fusions indicate that (i) translation of the carA gene can be initiated in vivo without an AUG codon but very likely with an UUG or an AUU codon; (ii) the carAB downstream promoter is repressed by arginine; and (iii) the carAB upstream promoter is repressed by pyrimidines and subject to stringent control. When carried by a multicopy plasmid the carAB control region escapes repression by arginine and pyrimidines. The existence of a pyrimidine repressor, present in limiting amounts in the cell, is therefore postulated.
Nucleotide sequence of a growth-related mRNA encoding a member of the prolactin-growth hormone family.
Authors: Linzer D I , Nathans D
Published: May 2006

As part of the proliferative response to serum, mouse 3T3 cells produce a set of growth-related mRNAs identified by hybridization to cloned cDNAs. One of these mRNAs, which is about 1 kilobase long, appears within a few hours after stimulation of resting cells with serum or platelet-derived growth factor and reaches a high level during the transition from the G1 to the S phase of growth. This mRNA is translated in vitro into a protein of approximately 25 kilodaltons. The corresponding cloned cDNA of 791 base pairs has been sequenced; it contains a single open reading frame that encodes a protein of 224 amino acids with extensive sequence homology to mammalian prolactins. The initial 29-amino acid segment of the encoded protein resembles the signal sequences of prehormones. That the growth-related protein is not mouse prolactin is indicated by comparison of its predicted amino acid composition with that of mouse prolactin and by the distinct fragment patterns seen when restricted mouse DNA is probed with the cloned cDNA or rat prolactin cDNA. Therefore, the growth-related protein appears to be a new member of the prolactin-growth hormone family. Because of its relationship to prolactin and growth hormone and its association with cell proliferation, the protein has been called "proliferin."
Molecular cloning of human terminal deoxynucleotidyltransferase.
Authors: Peterson R C , Cheung L C , Mattaliano R J , Chang L M , Bollum F J
Published: May 2006

A cDNA of the human terminal deoxynucleotidyltransferase (TdT; "terminal transferase," EC 2.7.7.31) was isolated from a human lymphoblastoid cell cDNA library in lambda gt 11 by using immunological procedures. Four inserts containing 723 to 939 base pairs were recloned in pBR322 for hybridization and preliminary sequence studies. mRNA selected by hybridization to recombinant DNA was translated to a 58-kDa peptide that specifically immunoprecipitated with rabbit antibodies to calf terminal transferase and mouse monoclonal antibody to human terminal transferase. Blot hybridization of total poly(A)+ RNA from KM3 (TdT+) cells with nick-translated pBR322 recombinant DNA detected a message of about 2000 nucleotides, sufficient to code for the 580 amino acids in the protein. mRNA from terminal transferase- cells gave no signal in hybrid selection or RNA blot hybridization. The complete sequence of the 939-base-pair insert sequence was obtained from deletions cloned in pUC8. The DNA sequence contains an open reading frame coding for 238 amino acids, about 40% of the protein. Three peptides isolated by HPLC from tryptic digests of succinylated 58-kDa calf thymus terminal transferase were sequenced, providing 20, 18, and 22 residues of peptide sequence. A search of the translated sequence of the 939-base-pair insert shows three regions beginning after arginine that have greater than 90% homology with the sequence determined from the calf thymus terminal transferase peptides. These results provide unambiguous evidence that the human terminal transferase sequence has been cloned.
Transformation-associated proteins in murine B-cell lymphomas that are distinct from Abelson virus gene products.
Authors: Pillemer E , Whitlock C , Weissman I L
Published: May 2006

In an effort to identify cellular proteins that may be involved in the Abelson murine leukemia virus (A-MuLV) transformation process, we have isolated a hybridoma antibody (6C3) that detects a tumor-associated antigen in all A-MuLV-induced pre-B-cell lymphomas. The 6C3 antibody immunoprecipitates two molecules of Mr 160,000 and Mr 125,000 from metabolically labeled A-MuLV tumors. The two proteins recognized by the 6C3 antibody are distinct from the A-MuLV-transforming protein in that they lack viral gag determinants and are neither phosphoproteins nor protein kinases. The 6C3 proteins can be detected in all A-MuLV pre-B-cell lymphomas and some nonviral B lymphomas but are not detected on any other tumor or normal cell, including A-MuLV-transformed fibroblast lines. Thus, the 6C3 proteins may represent the products of novel cellular genes whose expression is induced, stabilized, or amplified in B-cell tumors of both viral and nonviral origin. Further evidence in support of this hypothesis is provided by the finding that 6C3 antigen expression correlates with autonomous cell growth and the transformed phenotype in both normal bone marrow cultures and those infected with A-MuLV.
Distribution of DNA insertion element IS5 in natural isolates of Escherichia coli.
Authors: Green L , Miller R D , Dykhuizen D E , Hartl D L
Published: May 2006

DNA from Escherichia coli strains in a reference collection of 72 recent natural isolates (ECOR strains) and 25 natural isolates from the "pre-antibiotic" period 1930-1940 (Murray strains) were studied to determine the genomic abundance of insertion element IS5 and the size of genomic restriction fragments carrying sequences homologous to IS5. Among the ECOR strains, nearly two-thirds lack DNA sequences that hybridize with IS5, and one-half of the remainder have only one copy. Among strains in which IS5 is present, extensive variation in the size of IS5-bearing restriction fragments occurs, in many cases allowing distinction among strains that are judged to be nearly identical in genotype because of the identical electrophoretic mobility of the enzyme coded by each of 11 chromosomal loci. Among the Murray strains in which IS5 is present, the average number of elements per strain is larger, but not markedly so, than among recent isolates. Comparison between duplicate strains in the Murray collection suggests that the rate of accumulation of IS5 elements in prolonged storage in stab tubes corresponds to an apparent probability of transposition of approximately 0.008 +/- 0.002 per IS5 element per year. Because of the extensive genetic variation among strains, insertion elements such as IS5 would seem to be convenient genetic markers with which to detect recent common ancestry among strains.
Genomes of evolutionarily divergent members of the human T-cell leukemia virus family (HTLV-I and HTLV-II) are highly conserved, especially in pX.
Authors: Shaw G M , Gonda M A , Flickinger G H , Hahn B H , Gallo R C , Wong-Staal F
Published: May 2006

Human T-cell leukemia virus (HTLV) is a family of related human T-lymphotropic retroviruses closely linked with certain human T-cell malignancies and associated with many cases of acquired immunodeficiency syndrome (AIDS). We isolated and molecularly cloned HTLV from patients with both types of clinical disorders and found by restriction endonuclease mapping and core and envelope protein analysis that at least two evolutionarily divergent viral subgroups exist, HTLV-I and HTLV-II. Previous studies have failed to detect significant nucleotide sequence homology between HTLV-I and HTLV-II even though these different members of the HTLV family share certain biologic properties such as T-cell tropism and transformation. To further test these viruses for conserved regions in their genomes, we examined hybridization between HTLV-I and HTLV-II by using Southern blotting and heteroduplex mapping at different melting points. These two techniques produced similar results, showing that HTLV-I and HTLV-II proviruses have, in fact, strongly conserved nucleotide sequences in the pX region and lesser although still substantial homology in the LTR, gag, pol, and env regions. These data provide experimental evidence that HTLV-II, like HTLV-I, contains pX sequences. Although the function of pX is unknown, its conservation in evolutionarily divergent human T-lymphotropic viruses implies a biologically important function. It is possible, but unproven, that pX could encode proteins involved in T-cell tropism, cell transformation, immune suppression, or other biologic actions characteristic of the HTLV family.
Interactions of maitotoxin with voltage-sensitive calcium channels in cultured neuronal cells.
Authors: Freedman S B , Miller R J , Miller D M , Tindall D R
Published: May 2006

The dinoflagellate toxin maitotoxin (MTX) stimulated 45Ca2+ uptake in cultured NG108-15 neuroblastoma X glioma cells. Depolarizing stimuli (e.g., 50 mM K+) produced an immediate stimulation in Ca2+ uptake, whereas that produced by MTX occurred only after a lag period of about 2 min. MTX did not stimulate Ca2+ uptake into fibroblasts. Both 50 mM K+- and MTX-stimulated Ca2+ uptake was blocked by organic calcium channel antagonists (nitrendipine, D-600, diltiazem) at very low concentrations. Cd2+ was also a potent blocker. The novel dihydropyridine BAY K8644 enhanced Ca2+ uptake in the presence of 50 mM K+ but had no effect in 5 mM Ca2+. However, in the presence of MTX, BAY K8644 stimulated Ca2+ uptake in 5 mM K+. The effects of MTX were not blocked by tetrodotoxin but were decreased in Na+-free medium. MTX did not stimulate Na+ uptake into NG108-15 cells and did not alter [3H]nitrendipine binding to rat brain cortical synaptosomes. It is concluded that MTX may alter the voltage dependence of calcium-channel activation.
Molecular analysis of the recF gene of Escherichia coli.
Authors: Blanar M A , Sandler S J , Armengod M E , Ream L W , Clark A J
Published: May 2006

We analyzed the nucleotide sequence of a 1.325-kilobase region of wild-type Escherichia coli containing a functional recF gene and six Tn3 mutations that inactivate recF. The analysis shows a potentially translatable reading frame of 1071 nucleotides, which is interrupted by all six insertions. A protein of 40.5 kilodaltons would result from translation of the open reading frame, and a radioactive band of protein of an apparent molecular weight of approximately 40 kilodaltons was seen by the maxicell method using a recF+ plasmid. Putative truncated peptides were seen when two recF::Tn3 mutant plasmids were used. Differential expression of dnaN and recF from a common promoter was noted. recF332::Tn3 was transferred to the chromosome where, in hemizygous condition, it produced UV sensitivity indistinguishable from that produced by two presumed recF point mutations.
Bovine leukemia virus: unique structural features of its long terminal repeats and its evolutionary relationship to human T-cell leukemia virus.
Authors: Sagata N , Yasunaga T , Ogawa Y , Tsuzuku-Kawamura J , Ikawa Y
Published: May 2006

The nucleotide sequence of the long terminal repeat (LTR) of bovine leukemia virus, a unique oncogenic retrovirus of cattle, was determined. The LTR consisted of 530 base pairs (bp) with an inverted repeat of 6 bp at its 5' and 3' ends, flanked by a direct repeat of 6 bp of host cell origin. A tRNAPro binding site for minus-strand DNA synthesis followed the 5' LTR. The U3 region contained putative transcriptional promoters, "CAT" box and "TATA" box, but they had peculiar sequences (C-C-A-A-C-T and G-A-T-A-A-A-T). The U3 region also contained a potential enhancer element, whose sequence partially resembled those of other viral and cellular, especially of immunoglobulin, enhancers. The most striking structural feature of the LTR was an exceptionally long R region (228 bp), which separated a poly(A) addition signal (A-A-T-A-A-A) from a poly(A) site as far apart as 260 bp. The long R region was suggested to form a large stable hairpin structure on a nascent RNA chain, making the two transcription termination signals close together and thus ensuring normal termination of the chain. This structural feature of the bovine leukemia virus LTR was analogous to that of human T-cell leukemia virus LTR and, in fact, slight sequence homology (at most 50%) was observed between the R regions of these two retroviruses, indicating their evolutionary relationship. The unique structural feature of bovine leukemia virus and human T-cell leukemia virus LTRs may thus bear some relation to the biological features commonly shared by these retroviruses.
Differential DNase I sensitivity of the albumin and alpha-fetoprotein genes in chromatin from rat tissues and cell lines.
Authors: Nahon J L , Gal A , Erdos T , Sala-Trepat J M
Published: May 2006

We have examined the DNase I sensitivity of the albumin and alpha-fetoprotein (AFP) genes in different rat tissues (adult liver and kidney) and cloned cell lines (hepatoma 7777-C8, JF1 fibroblasts), which show drastic differences in the level of expression of these two genes. This was done by studying the disappearance of defined restriction endonuclease fragments of these genes as a function of limited DNase I digestion. The sensitivity of these genes was compared to that of a gene not expressed in the hepatic cells and to that of a ubiquitously expressed gene. In nuclei from adult rat liver the albumin and AFP genes were preferentially degraded by the nucleolytic action of DNase I, whereas they were not in rat kidney nuclei. In the hepatoma cells the AFP gene was much more sensitive to DNase I digestion than the albumin gene; both genes were very resistant to DNase I action in fibroblastic nuclei. When analyzed in relation to the level of gene expression our results indicate that alterations in the chromatin structure of the albumin and AFP genes might be involved in the early establishment of the tissue-specific potential of overt gene expression; such alterations reflected in an altered DNase I sensitivity do not appear to be responsible for the changes in gene activity occurring during the terminal differentiation of the hepatocyte; and modifications in the chromatin structure of these genes might occur during oncogenic events; these structural modifications could be related to the changes in gene expression observed in hepatocarcinogenic processes.
Statistical mechanics of the B----Z transition of DNA: contribution of diffuse ionic interactions.
Authors: Soumpasis D M
Published: May 2006

A theoretical framework is constructed to treat the effects of salt ions on polyionic structural transitions in the absence of specific ion binding. As an application, the salt concentration-dependent part of the free energy difference governing the B----ZI transition of [d(C-G) . d(C-G)]6 in 1:1 electrolytes is calculated; quantitative agreement with the experimental findings is obtained. The effects of temperature and multivalent cations are also discussed. Preliminary calculations indicate that the ZII conformation in solution is thermodynamically less favorable than ZI in the high-salt regime (2.0-5.0 M) but more favorable than ZI below 2.0 M salt.
Chromosomal organization of the human dihydrofolate reductase genes: dispersion, selective amplification, and a novel form of polymorphism.
Authors: Anagnou N P , O'Brien S J , Shimada T , Nash W G , Chen M J , Nienhuis A W
Published: May 2006

The human dihydrofolate reductase (DHFR; tetrahydrofolate dehydrogenase; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) gene family includes a functional gene (hDHFR) and at least four intronless genes. Three intronless genes (hDHFR-psi 2, hDHFR-psi 3, and hDHFR-psi 4) are identifiable as pseudogenes because of DNA sequence divergence from the functional gene with introns, while one intronless gene (hDHFR-psi 1) is completely homologous to the coding sequences of the functional gene. Analysis of genomic DNA from two panels of somatic human-rodent cell hybrids with specific molecular probes provide insight into the chromosomal organization and assignment of these genes. The five genes are dispersed in that each one is found on a different chromosome. The functional gene hDHFR has been assigned to chromosome 5, and one pseudogene (hDHFR-psi 4), to chromosome 3. In a human cell line (HeLa) that was selected for methotrexate resistance, the functional locus became amplified, while there was no amplification of the four intronless pseudogenes. hDHFR-psi 1 was found to be present in DNA of some individuals and absent from DNA of others, consistent with a recent evolutionary origin of this gene originally suggested by its sequence identity to the coding portions of the functional gene. The presence or absence of this intronless pseudogene represents a previously unreported form of DNA polymorphism.
Isolation of HLA locus-specific DNA probes from the 3'-untranslated region.
Authors: Koller B H , Sidwell B , DeMars R , Orr H T
Published: May 2006

When human class I cDNA clones containing coding sequences are used to probe genomic DNA, 15-20 fragments, each containing a complete class I gene or pseudogene, are seen. Identification of which genomic DNA segments encode the HLA-A and -B antigens has to date required transfection of mouse L cells with cloned class I genes or analysis of HLA loss mutants. In this report we show that under high-stringency conditions, probes constructed from the 3'-untranslated region can be used to specifically identify the segments of DNA that encode the HLA-A and -B antigens in the human lymphoblastoid cell line 721. Examination of DNA from unrelated individuals indicates that these probes are locus specific and will permit identification of HLA-A and -B genes in the population.
A molecular hybrid of the H-2Dd and H-2Ld genes expressed in the dm1 mutant.
Authors: Burnside S S , Hunt P , Ozato K , Sears D W
Published: May 2006

Sequential immunoprecipitates show that H-2dm1 mutant cells express a hybrid "H-2D/L" antigen exhibiting determinants normally associated with two different gene products of the parental d haplotype-i.e., the H-2Dd and H-2Ld antigens. The hybrid H-2D/Ldm1 antigen appears to consist of a portion of the NH2-terminal extracellular half of the H-2Dd antigen "fused" to a portion of the COOH-terminal extracellular half of the H-2Ld antigen. This structure is inferred from the reactivity of dm1 antigens with cytotoxic T lymphocytes specific for H-2Ld determinants and with monoclonal antibodies specific for determinants in the structural domains of H-2Ld or H-2Dd. The H-2D/Ldm1 molecule apparently retains all of the third external domain (C2 or alpha 3) and part of the second external domain (C1 or alpha 2) of H-2Ld, but its first external domain (N or alpha 1) derives from H-2Dd. From these findings and from previous peptide mapping studies, we propose that the H-2D/Ldm1 antigen is the product of a hybrid gene that has resulted from an unequal crossover between the parental H-2Dd and H-2Ld genes, leaving the N exon and part of the C1 exon of the H-2Dd gene joined to the H-2Ld gene beginning somewhere within its C1 exon.
Identity of hepatic membrane transport systems for bile salts, phalloidin, and antamanide by photoaffinity labeling.
Authors: Wieland T , Nassal M , Kramer W , Fricker G , Bickel U , Kurz G
Published: May 2006

Phalloidin, a bicyclic heptapeptide, and antamanide, a monocyclic decapeptide from the poisonous mushroom Amanita phalloides, interact with bile-salt-binding polypeptides of the hepatocyte membrane, as demonstrated by photoaffinity labeling using the photolabile bile salt derivative 7,7,-azo-3 alpha, 12 alpha-dihydroxy-5 beta-cholan-24-oic acid, either unconjugated or taurine conjugated. With the photolabile derivatives of phalloidin, N-delta-(4-[(1-azi-2,2,2-trifluoroethyl) benzoyl]-beta-alanyl)-delta-aminophalloin, (N epsilon-[4-(1-azi-2,2,2-trifluoroethyl)benzoyl]lys6)-anta manide, the same membrane polypeptides with apparent MrS of 54,000 and 48,000 were labeled as with the photolabile derivatives of unconjugated and conjugated bile salts. The presence of bile salts decreased markedly the extent of labeling of these phalloidin- and antamanide-binding polypeptides. These results indicate that hepatic uptake systems for bile salts, phallotoxins, and the cycloamanide antamanide are identical, thus explaining the organotropism of phallotoxins.
Post-classification version of Jordan's theorem on finite linear groups
Authors: Weisfeiler Boris
Published: May 2006

Using classification of finite simple groups, I show that a finite subgroup
G
of GL

n

(C), where C = the complex numbers, contains a commutative normal subgroup
M
of index at most (
n
+ 1)!

n
alogn+b

. Moreover, if
G
is primitive and does not contain normal subgroups that are direct products of large alternating groups, then the factor (
n
+ 1)! can be dropped. I further show that similar statements hold also in characteristics
p
≥ 2, if one takes
M
to be an extension of a group of Lie type of characteristic
p
by a solvable group that has a normal
p
-subgroup with commutative
p
′-quotient. These results improve the celebrated theorems of Jordan and of Brauer and Feit.
125I-labeled crosslinking reagent that is hydrophilic, photoactivatable, and cleavable through an azo linkage.
Authors: Denny J B , Blobel G
Published: May 2006

A radioactive crosslinking reagent, N-[4-(p-azido-m-[125I]iodophenylazo)benzoyl]-3-aminopropyl-N' -oxysulfosuccinimide ester, has been synthesized. The reagent is photoactivatable, water-soluble, cleavable through an azo linkage, and labeled with 125I at the carrier-free specific activity of 2000 Ci/mmol. Any protein derivatized with the reagent is thus converted into an 125I-labeled photoaffinity probe. Crosslinks are formed following photolysis with 366-nm light, and cleavage by sodium dithionite results in the donation of radioactivity to the distal partner in crosslinked complexes. The newly labeled proteins are then analyzed by gel electrophoresis and autoradiography. The compound was prepared by iodination of N-[4-(p-aminophenylazo)benzoyl]-3-aminopropionic acid using carrier-free Na125I and chloramine-T, followed by azide formation and conversion to the water-soluble sulfosuccinimide ester. As a model system, protein A-Sepharose was derivatized with the reagent under subdued light. Each derivatized protein A molecule contained only one crosslinker. The derivatized protein A-Sepharose was then photolyzed in the presence of human serum and subsequently treated with sodium dithionite. Analysis of the serum by gel electrophoresis revealed that 1.1% of the radioactive label originally present on the protein A-Sepharose was transferred to the heavy chain of IgG, which was the most intensely labeled protein in the gel. The next most intensely labeled protein was IgG light chain, which incorporated radioactivity that was lower by a factor of 3.6 than that of the heavy chain. These results demonstrated the specificity of the derivatized protein A-Sepharose as a photoaffinity probe. Photolabeling of IgG was the result of nitrene-mediated reactions and was not due to the incorporation of free 125I.
Vasopressin gene is expressed at low levels in the hypothalamus of the Brattleboro rat.
Authors: Majzoub J A , Pappey A , Burg R , Habener J F
Published: May 2006

The Brattleboro rat carries as a recessive trait the inability to synthesize hypothalamic vasopressin and its related neurophysin but is able to synthesize oxytocin and its neurophysin. Brattleboro rats homozygous for this trait have no immunologically detectable circulating vasopressin and manifest a complete syndrome of diabetes insipidus, which is corrected with vasopressin replacement therapy. Such a defect could be due to absence of the gene encoding vasopressin, the presence of an abnormal gene, or a variety of transcriptional or posttranscriptional abnormalities. We report here that the hypothalamus of the Brattleboro rat contains detectable, although markedly reduced, levels of an mRNA indistinguishable in size with and similar in sequence to authentic vasopressin mRNA. Corresponding levels of oxytocin mRNA were the same in Brattleboro and normal rat hypothalami. These findings indicate that the Brattleboro rat expresses a vasopressin gene, but at a reduced level.
Neoplastic human fibroblast proteins are related to epidermal growth factor precursor.
Authors: Burbeck S , Latter G , Metz E , Leavitt J
Published: May 2006

We report the amino acid composition of two polypeptides, p788 and p789. These polypeptides are reliable markers for neoplastic transformation in human fibroblasts. Their compositions are unusually rich in cysteine and serine. Because the recently reported amino acid sequence of mouse epidermal growth factor precursor (prepro-EGF) is also rich in those two amino acids and because the role of p788 and p789 as markers for neoplastic transformation is consistent with the fact that epidermal growth factor has been shown to play some role in transformation, we investigated the hypothesis that p788 and p789 are related to prepro-EGF. We compared the amino acid composition of p788 with that of all possible interior domains of prepro-EGF of appropriate length. We found that the composition of p788 is remarkably similar to that of residues 630-880 of prepro-EGF. The similarity is sufficiently strong to support the conclusion that it reflects amino acid sequence homology.
Replication of bacteriophage phi 29 DNA in vitro: the roles of terminal protein and DNA polymerase.
Authors: Watabe K , Leusch M , Ito J
Published: May 2006

phi 29 DNA replication is initiated by the formation of a covalent complex between the viral-coded terminal protein and dAMP (TP-dAMP). This initiation reaction system has been reconstituted from two phage-encoded proteins, the terminal protein and DNA polymerase. The phi 29 DNA polymerase was purified from phage-infected cells by using poly(dA) X p(dT)12-18 as an assay template. The purified polymerase has an apparent molecular mass of 68 kDa in its native form and it appears to function as a monomer. The terminal protein was purified to homogeneity from Escherichia coli cells harboring a cloned plasmid that contained a phi 29 gene 3 segment. The molecular mass of the purified terminal protein was about 30 kDa in both the denatured and the native form. The protein apparently functions as a monomer. When the terminal protein and DNA polymerase were incubated in the presence of dATP, Mg2+, and phi 29 DNA-protein as template, the terminal protein bound covalently to dAMP. This reaction did not require ATP. In addition, these two purified fractions catalyzed DNA chain elongation from both ends of phi 29 DNA, yielding the expected 9- to 12-base fragment when assayed in the presence of 2',3'-dideoxycytidine triphosphate. These results indicate that phi 29 DNA polymerase catalyzes formation of the terminal protein-dAMP complex and can also catalyze chain elongation at least 9-12 bases from both ends of phi 29 DNA.
The increase in intracellular free calcium associated with IgG gamma 2b/gamma 1 Fc receptor-ligand interactions: role in phagocytosis.
Authors: Young J D , Ko S S , Cohn Z A
Published: May 2006

The concentration of cytosolic free calcium, [Ca2+]i, was measured in J774 mouse macrophages by use of the fluorescent indicator quin-2. Resting [Ca2+]i was 87 nM. Addition of a number of specific ligands to the immunoglobulin gamma 2b/gamma 1 Fc receptor resulted in a transient increase in [Ca2+]i, the magnitude of which depended on the extent of receptor aggregation. Monovalent ligands gave only a small Ca2+ signal but blocked cell response to subsequent addition of multivalent ligands. Incubation with antibody-coated erythrocytes raised macrophage [Ca2+]i to micromolar levels. [Ca2+]i changes were only partially inhibited by the absence of external Ca2+, suggesting the release of Ca2+ from internal stores in addition to an influx of external Ca2+. These internal stores were not limited to mitochondria. An optimal range of [Ca2+]i was required for phagocytosis. Buffering [Ca2+]i with quin-2 and treating cells with quinine in the absence of external Ca2+ resulted in inhibition of phagocytosis. Increasing [Ca2+]i to micromolar levels with the calcium ionophore A23187 also resulted in similar inhibitory effects. We suggest the involvement of localized cytosolic Ca2+ gradients in generating the signals necessary for phagocytosis.
Simultaneous in situ detection of viral RNA and antigens.
Authors: Brahic M , Haase A T , Cash E
Published: May 2006

We have combined in situ hybridization and immunocytochemistry to detect RNA and proteins in the same cell. We envision a wide range of applications for this method, and, as one example, we show that viral genomes and capsid antigens can be detected simultaneously in paraffin sections of the central nervous system of mice infected with Theiler's virus, the causative agent of a slow demyelinating disease.
Receptor-mediated endocytosis of low density lipoprotein: somatic cell mutants define multiple genes required for expression of surface-receptor activity.
Authors: Kingsley D M , Krieger M
Published: May 2006

We have used cell fusion and mutant reversion analysis to study a collection of Chinese hamster ovary (CHO) cell mutants that are unable to bind and internalize low density lipoprotein (LDL). Pairwise cell fusions show that these LDL receptor-deficient mutants fall into three recessive complementation groups, ldlA, ldlB, and ldlC. Complementation was detected by observing the uptake of fluorescent LDL and was quantitated by measuring the degradation of 125I-labeled LDL by isolated hybrid cells. Previous studies had defined a fourth recessive complementation group, ldlD. Complementation tests between CHO cells and human fibroblasts suggested that the defects in mutants of the ldlA complementation group are analogous to those in a patient with homozygous familial hypercholesterolemia. A revertant of an ldlA mutant was isolated and appeared to be heterozygous at the ldlA locus. The phenotype of this revertant was similar to that of cells from patients with the heterozygous form of familial hypercholesterolemia. Together with recent DNA transfection studies, these results suggest that the ldlA locus is the structural gene for the LDL receptor in CHO cells. Mutants in the ldlB, ldlC, and ldlD complementation groups must have defects in genes that are required for either the regulation, synthesis, transport, recycling, or turnover of LDL receptors.
Liposomes as targets for granule cytolysin from cytotoxic large granular lymphocyte tumors.
Authors: Blumenthal R , Millard P J , Henkart M P , Reynolds C W , Henkart P A
Published: May 2006

Purified cytoplasmic granules from rat large granular lymphocyte tumors having natural killer activity and/or antibody-dependent cell-mediated cytotoxicity induced a rapid, dose-dependent release of the water-soluble marker carboxyfluorescein from liposomes made of phosphatidylcholine. A solubilized, partially purified cytolytic preparation termed "cytolysin" from these granules showed identical properties. Marker release induced by granules or the cytolysin was strongly dependent on the presence of Ca2+ at a concentration of 0.1 mM or higher in the medium; Ca2+ could be replaced by higher concentration of Sr2+ but not by Ba2+ or by Mg2+. These properties strikingly parallel the lytic effects that granules and granule cytolysin exert on cells. Marker release from liposomes was stopped instantaneously when an excess of EGTA was added to the medium. The remaining carboxyfluorescein inside the liposomes was present at the original internal concentration, indicating that marker release was all-or-none from individual liposomes. Liposomes comprised of lipid in the solid phase released marker more slowly than did comparable liposomes containing fluid-phase lipids. Variation of the lipid headgroup had only minor effects on the cytolysin-induced marker release. Electron microscopy of liposomes exposed to cytolysin in the presence of Ca2+ showed cylindrical structures of 15-nm diameter inserted into the membrane concomitant with the penetration of negative stain into the liposome. These properties of large granular lymphocyte granule cytolysin strongly suggest that it operates through a mechanism similar to the membrane attack of complement.
Activity and the control of ganglion cell death in the rat retina.
Authors: Fawcett J W , O'Leary D D , Cowan W M
Published: May 2006

In newborn rats each retina projects principally to the contralateral superior colliculus, but there is also a sparse projection to the whole of the ipsilateral superior colliculus. During the first 2 weeks postnatally the ipsilateral projection normally becomes restricted to the rostromedial part of the superior colliculus. The restriction of this projection is due to the preferential death of ipsilaterally projecting retinal ganglion cells and is apparently the result of competition between optic fibers from the two eyes, since it can be prevented by enucleation of the opposite eye at birth. To determine if electrical activity plays a role in the normal restriction of the ipsilateral retinocollicular projection, the sodium channel-blocking agent tetrodotoxin was administered to one or both eyes during the first 2 weeks postnatally. Tetrodotoxin blockade of activity in one eye resulted in the persistence of a sparse projection from the opposite eye throughout the ipsilateral superior colliculus and the survival of a substantial number of the ipsilaterally projecting retinal ganglion cells in that eye that would normally have died. When both eyes were treated with tetrodotoxin no restriction of the ipsilateral projection was seen on either side. These findings suggest that the competition between retinal ganglion cell axons (either for terminal space or an essential trophic factor), which normally leads to retinal ganglion cell death and the restriction of the ipsilateral retinocollicular projection, is mediated in some way by electrical activity.
Adenosine strongly potentiates pressor responses to nicotine in rats.
Authors: von Borstel R W , Renshaw A A , Wurtman R J
Published: May 2006

Intravenous infusion of subhypotensive doses of adenosine strongly potentiates the pressor response of anesthetized rats to nicotine. A dose of nicotine (40 micrograms/kg, i.v.), which, given alone, elicits a peak increase in diastolic pressure of approximately equal to 15 mm Hg, increases pressure by approximately equal to 70 mm Hg when arterial plasma adenosine levels have been increased to 2 microM from a basal concentration of approximately equal to 1 microM. The pressor response to cigarette smoke applied to the lungs is also strongly potentiated during infusion of adenosine. Slightly higher adenosine concentrations (approximately equal to 4 microM) attenuate pressor responses to electrical stimulation of preganglionic sympathetic nerves, or to injections of the alpha-adrenergic agonist phenylephrine, but continue to potentiate pressor responses to nicotine. Low doses (0.25-5 micrograms/kg) of the synthetic adenosine receptor agonists 5'-N-cyclopropylcarboxamidoadenosine, 2-chloroadenosine, and N6-L-phenylisopropyladenosine also potentiate pressor responses to nicotine. Caffeine and theophylline (10 mg/kg) block the potentiating effect of adenosine, and also decrease basal responses to nicotine, suggesting that endogenous adenosine might normally potentiate some nicotine responses. The synergism between nicotine and adenosine appears to take place within sympathetic ganglia.
Mapping of N6-methyladenosine residues in bovine prolactin mRNA.
Authors: Horowitz S , Horowitz A , Nilsen T W , Munns T W , Rottman F M
Published: May 2006

N6-Methyladenosine (m6A) residues, which are found internally in viral and cellular mRNA populations at the sequences Apm6ApC and Gpm6ApC, have been proposed to play a role in mRNA processing and transport. We have developed a sensitive approach to analyze the level and location of m6A in specific purified cellular mRNAs in an attempt to correlate m6A location with function. Polyadenylylated mRNA is hybridized to cDNA clones representing the full size mRNA under study or fragments of it, and the protected RNA is digested and labeled with polynucleotide kinase in vitro. After enrichment for m6A with anti-m6A antibody, the [32P]-pm6A is separated on TLC plates, and compared with the total amount of radiolabeled nucleotides. Using this combination of in vitro RNA labeling and antibody selection, we were able to detect m6A in purified stable mRNAs that cannot be readily labeled in cells with greater sensitivity than was possible by previous techniques. We applied this technique to bovine prolactin mRNA and showed that this mRNA contains m6A. Moreover, all of the m6A residues in this message are found within the 3' two-thirds of the molecule and are highly concentrated (61%) within a sequence of 108 nucleotides at the 3' noncoding region of the message. The nonrandom distribution of m6A in a specific cellular mRNA, as demonstrated for bovine prolactin, will have to be taken into account when designing a model for m6A function.
Cloning of terminal transferase cDNA by antibody screening.
Authors: Landau N R , St John T P , Weissman I L , Wolf S C , Silverstone A E , Baltimore D
Published: May 2006

A cDNA library was prepared from a terminal deoxynucleotidyltransferase-containing thymoma in the lambda phage vector lambda gt11. By screening plaques with anti-terminal transferase antibody, positive clones were identified of which some had beta-galactosidase-cDNA fusion proteins identifiable after electrophoretic fractionation by immunoblotting with anti-terminal transferase antibody. The predominant class of cross-hybridizing clones was determined to represent cDNA for terminal transferase by showing that one representative clone hybridized to a 2200-nucleotide mRNA in close-matched enzyme-positive but not to enzyme-negative cells and that the cDNA selected a mRNA that translated to give a protein of the size and antigenic characteristics of terminal transferase. Only a small amount of genomic DNA hybridized to the longest available clone, indicating that the sequence is virtually unique in the mouse genome.
Specific membrane receptors for atrial natriuretic factor in renal and vascular tissues.
Authors: Napier M A , Vandlen R L , Albers-Schönberg G , Nutt R F , Brady S , Lyle T , Winquist R , Faison E P , Heinel L A , Blaine E H
Published: May 2006

Membranes from rabbit aorta and from rabbit and rat kidney cortex possess high-affinity (Kd = 10(-10) M) specific binding sites for atrial natriuretic factor (ANF). Similar high-affinity sites are present in an established cell line from pig kidney, LLC-PK1. Results of fractionation studies indicate that the receptors are localized in the plasma membrane of these tissues. The binding is time-dependent and saturable. An excellent quantitative correlation was found between the affinity of synthetic ANF and analogs of intermediate activity to aorta membranes and the half-maximal concentration needed for relaxation of rabbit aorta rings contracted by addition of serotonin. Furthermore, the binding affinity of the receptor in kidney membranes is consistent with the concentration required for in vivo natriuresis in the rat. Biologically inactive synthetic ANF fragments and other peptide hormones such as angiotensin II and vasopressin do not significantly inhibit binding. These data suggest that the receptors for ANF in vascular and renal tissues are responsible for mediating the physiological actions of this peptide in these target tissues.