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S M Weissman

Publications and source records attributed to S M Weissman.

At least 163 records · Page 9Linked to original sources

Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer.

We have isolated a cDNA clone for one of the HLA-B locus alloantigens by hybridization with a 30-nucleotide-long DNA probe. The probe was isolated from a reverse transcriptase (RNA-dependent DNA nucleotidyltransferase)-catalyzed cDNA synthesis reaction on poly(A)-mRNA in which an oligonucleotide (5'-32P)dC-T-T-C-T-C-C-A-C-A-TOH served as a primer and in which dideoxynucleoside triphosphates were used to reduce the size and heterogeneity of the cDNA products. The desired cDNA clone was isolated from a library of recombinant cDNA clones in the plasmid pBR322. The partial nucleotide sequence of the cDNA clone corresponds to the amino acid sequence of HLA-B7 antigen. The approach described in this paper is extremely sensitive and may be useful in cloning other genes for which the corresponding mRNA is present at low levels. This cDNA clone is nearly full length and can be used to isolate and to study the genes within the HLA region and to obtain expression of HLA-B peptides in cells.

Base Sequence↗

Unspliced functional late 19S mRNAs containing intervening sequences are produced by a late leader mutant of simian virus 40.

dl-805 is a viable deletion mutant of simian virus 40 lacking 187 of the 202 nucleotides coding for the principal leader of the wild-type viral late mRNAs. The structures of the mutant late 16S and 19S RNAs and proteins were examined with the following principal findings. (i) The vast majority, if not all, of the 19S RNAs are unspliced over their entire length. These RNAs contain intervening sequences and serve as mRNAs. The vast majority, if not all, of the 16S RNAs, however, retain the characteristic wild-type splice. (ii) Mutant 16S and 19S RNAs do not terminate at residue 243 (located six nucleotides upstream from the 5' end of the deleted DNA segment), the principal 5' terminus of the wild-type late mRNAs; rather, they begin at a series of sites, mostly upstream, used infrequently or not at all by wild-type virus. (iii) Despite the presence of altered late mRNAs, dl-805 late proteins have electrophoretic mobilities similar to those of wild-type late proteins, and mutant virions contain the same relative quantities of late proteins as wild-type virions.

Base Sequence↗

Base substitution in an intervening sequence of a beta+-thalassemic human globin gene.

beta globin gene fragments from a patient with homozygous beta+-thalassemia have been cloned and subjected to restriction endonuclease, nucleotide sequence, and in vitro trancription analyses. Restriction endonuclease mapping of the cloned gene fragments revealed no deletions or other rearrangements, and transcription of the thalassemic gene appeared to be normal in vitro. However, nucleotide sequence analysis of the beta+-thalassemic gene fragments permitted identification of a single base change in the body of the small intervening sequence. This nucleotide change creates a sequence much like that of the 3' splice site of the small intervening sequence. The presence of a potential anomalous splicing site as a result of this base change suggests a mechanism for defective posttranscriptional processing of beta globin mRNA precursor molecules in beta+-thalassemia.

Base Sequence↗

Nucleotide sequence of the coding portion of human alpha globin messenger RNA.

The nucleotide sequence of the coding portion of human alpha globin mRNA has been determined by sequence analysis using human alpha globin cDNA cloned in bacterial plasmids. The sequence was obtained by a combination of direct sequence analysis of the cloned cDNA and analysis of cDNA obtained by primer extension, using short restriction endonuclease fragments of cloned alpha cDNA that were hybridized to human globin mRNA and elongated on the mRNA template by viral reverse transcriptase. The human alpha globin mRNA has an unexpectedly high G + C base composition (64.7%), similar to that observed for rabbit globin alpha mRNA, and displays a striking bias in the use of synonym codons for various amino acids. The bias in codon usage of human alpha globin mRNA is similar, with some exceptions, to that previously observed for rabbit alpha globin mRNA as well as for human and rabbit beta globin mRNAs. A detailed restriction endonuclease map of the human alpha globin cDNA is presented.

Amino Acid Sequence↗

Complete nucleotide sequence of the human delta-globin gene.

We present the complete nucleotide sequence of the human delta-globin gene. The general DNA sequence organization of the delta-globin gene is similar to that of other known globin genes. We have been unable to identify unambiguously the structural basis of the low level of expression characteristic of the delta-globin gene.

Amino Acid Sequence↗

The structure and evolution of the human beta-globin gene family.

We present the results of a detailed comparison of the primary structure of human beta-like globin genes and their flanking sequences. Among the sequences located 5' to these genes are two highly conserved regions which include the sequences ATA and CCAAT located 31 +/- 1 and 77 +/- 10 bp, respectively, 5' to the mRNA capping site. Similar sequences are found in the corresponding locations in most other eucaryotic structural genes. Calculation of the divergence times of individual beta-like globin gene pairs provides the first description of the evolutionary relationships within a gene family based entirely on direct nucleotide sequence comparisons. In addition, the evolutionary relationship of the embryonic epsilon-globin gene to the other human beta-like globin genes is defined for the first time. Finally, we describe a model for the involvement of short direct repeat sequences in the generation of deletions in the noncoding and coding regions of beta-like globin genes during evolution.

Animals↗

Nucleotide sequence of human G gamma globin messenger RNA.

The nucleotide sequences of the entire coding portion and 3'-untranslated portion of the human G gamma globin mRNA have been determined by direct nucleotide sequence analysis of cloned human G gamma globin cDNAs. These nucleotides sequences have been compared to the previously published sequences of the human beta globin mRNA and A gamma globin mRNA. No differences were detected between the coding sequences of G gamma and A gamma globin mRNAs except for the expected difference at codon No. 136, but a number of differences were detected in the 3'-untranslated sequences of the two mRNAs. Codon usage of G gamma globin mRNA, although similar to that of the related beta globin mRNA, displays a few notable differences such as the use of UUG for leucine and GGA for glycine that are not utilized in beta globin mRNA.

Base Sequence↗

Ubiquitous, interspersed repeated sequences in mammalian genomes.

DNA base sequence comparisons demonstrate that the principal family of 300-nucleotide interspersed human DNA sequences, the repetitive double-strand regions of HeLa cell heterogeneous nuclear RNA, and specific RNA polymerase III in vitro transcripts of cloned human DNA sequences are all representatives of a closely related family of sequences. A segment of approximately 30 residues of these sequences is highly conserved in mammalian evolution because it is also present in the interspersed repeated DNA sequences of Chinese hamsters. Further DNA sequence comparisons demonstrate that a portion of this highly conserved segment of repetitive mamalian DNA sequence is similar to a sequence found within a low molecular weight RNA that hydrogen-bonds to poly(A)-terminated RNA molecules of Chinese hamsters and a sequence that forms half of a perfect inverted repeat near the origin of DNA replication in papovaviruses.

Animals↗

RNA polymerase III transcriptional units are interspersed among human non-alpha-globin genes.

Cloned human DNA fragments containing globin genes are transcribed in vitro to form discrete RNA species. One transcription unit is located approximately 1500 base pairs upstream from the G-gamma-globin gene. This transcript is partially homologous to a polymerase III template located approximately 1000 base pairs upstream from the delta-globin gene and to DNA located a short distance downstream from the beta-globin gene.

Cloning, Molecular↗

Simian virus 40 early mRNA's. I. Genomic localization of 3' and 5' termini and two major splices in mRNA from transformed and lytically infected cells.

We have studied the structure of polyadenylated virus-specific cytoplasmic mRNA's in mouse and human cells transformed by simian virus 40 and in monkey cells infected with simian virus 40 in the presence of cytosine arabinoside by means of reverse transcriptase-catalyzed complementary DNA synthesis and complementary DNA sequencing. Abundant mRNA species containing splices from residues 4490 to 4557 (0.533 to 0.546 map units [m.u.]) and 4490 to 4837 (0.533 to 0.600 m.u.) were identified in both transformed and infected cells. Two principal reverse transcriptase stops were observed at the 5' termini of these mRNA's, both occurring with approximately equal frequency. The most distal of these stops was localized at residues 5152 to 5154 (0.660 m.u.), and the second was at residues 5147 to 5148 (0.659 m.u.). Several additional minor stops, between approximately 0.62 and 0.65 m.u., were also found on complementary DNA copied from transformed cell mRNA; in contrast, only one additional stop was present on complementary DNA copied from early lytic mRNA. These data suggest the presence of a prinicipal 5' terminus of early lytic and transformed cell mRNA's at residues 5152 to 5154 and raise the possibility of additional 5' termini at one or more locations in the 0.62 to 0.659 m.u. region of these mRNA's. Transformed cell mRNA was also found to contain a single 3' terminus at positions 2504 and 2505 (0.153 m.u.); termini lying beyond this site were not detected.

Animals↗