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Biomedical subjects

T Shenk

Publications and source records attributed to T Shenk.

At least 145 records · Page 8Linked to original sources
PubMed1984

Deletion of the gene encoding the adenovirus 5 early region 1b 21,000-molecular-weight polypeptide leads to degradation of viral and host cell DNA.

The adenovirus 5 mutant H5dl337 lacks 146 base pairs within early region 1B. The deletion removes a portion of the region encoding the E1B 21,000-molecular-weight (21K) polypeptide, but does not disturb the E1B-55K/17K coding region. The virus is slightly defective for growth in cultured HeLa cells, in which its final yield is reduced ca. 10-fold compared with wild-type virus. The mutant displays a striking phenotype in HeLa cells. The onset of cytopathic effect is dramatically accelerated, and both host cell and viral DNAs are extensively degraded late after infection. This defect has been described previously for a variety of adenovirus mutants and has been termed a cytocidal (cyt) phenotype. H5dl337 serves to map this defect to the loss of E1B-21K polypeptide function. In addition to its defect in the productive growth cycle, H5dl337 is unable to transform rat cells at normal efficiency.

Adenoviruses, Human↗
PubMed1983

Functional analysis of the nucleotide sequence surrounding the cap site for adenovirus type 5 region E1A messenger RNAs.

We have constructed a set of small, dispersed deletion mutations in the sequences surrounding the cap site of the adenovirus type 5 region E1A transcription unit. The effects of the mutations on E1A transcription were studied in vitro using a HeLa cell-free extract, and in vivo by reconstructing the mutations back into intact viral chromosomes and analyzing E1A messenger RNAs synthesized after infection of HeLa cells. The sequence between -35 and +20 (relative to the cap site at +1) was important for efficient E1A transcription and cap site selection in vitro. This region includes the "TATA" homology, which appeared essential for transcription. Sequences upstream of -35 were dispensable for transcription in vitro. Different results were found upon analysis of the same set of deletions in vivo. None of the mutations affected the steady-state levels of cytoplasmic, E1A-specific mRNAs found in infected HeLa cells by more than twofold. Deletions of the TATA homology, however, generated E1A mRNAs with heterogeneous 5' ends, and deletions downstream of the homology displaced the 5' end of mRNAs by about the size of the deletion.

Adenoviridae↗
PubMed1983

The adenovirus type 5 E1A transcriptional control region contains a duplicated enhancer element.

The adenovirus type 5 E1A transcriptional control region contains an element with enhancer properties located between -141 and -305 relative to the E1A cap site at +1. The enhancer element is located at or very close to a sequence required in cis for packaging of viral DNA. Deletion of the element reduces both the rate of transcription and steady-state levels of E1A mRNAs in virus-infected cells. Such deletions also cause a modest reduction in activity of the E1B transcription unit, which is located immediately downstream of the E1A unit. The enhancer element includes repeated core sequences. Function is retained when the element is moved to the 3' side of the E1A gene in either possible orientation, and the E1A enhancer functions in cis to enhance transformation by the herpesvirus thymidine kinase gene in both mouse and human cells.

Adenoviruses, Human↗
PubMed1982

Adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection.

Two adenovirus type 5 mutants were constructed to probe the function of the virus-encoded RNA polymerase III transcripts (VA RNAs). Each mutant fails to synthesize one of the two VA RNA species. The variant that does not produce the minor VAII species grows normally. The mutant that cannot synthesize the major VAI species grows more poorly than its parent. Analysis of the mutant's growth defect indicates that the adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection.

Adenoviruses, Human↗
PubMed1982

Fragments of the simian virus 40 transforming gene facilitate transformation of rat embryo cells.

Segments of the simian virus 40 (SV40) genome that encode only fragments of large tumor antigen can facilitate immortalization of secondary rat embryo cells. The phenotypes of the immortalized cells range from nearly "normal" to fully transformed. All of the cell lines contain SV40 sequences and express unstable NH2-terminal fragments of large tumor antigen. SV40 small tumor antigen does not appear to be essential for either immortalization or transformation.

Animals↗
PubMed1981

The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs.

We have observed three effects of deletion mutations on polyadenylation of late SV40 mRNAs. The first class of mutants lack segments (-3 to -14 bp) between the 5-AAUAAA-3' and normal poly(A) site. These mutants produce mRNas polyadenylated at new sites, downstream from the wild-type site. The poly(A) site is moved farther downstream as the deletions become larger; as a result, polyadenylation always occurs within an 11-19 nucleotide range from the AAUAAA sequence. The second class of mutants lack segments (-12 to -30 bp) between the AAUAAA sequence and the coding region of the mRNA. The poly(A) site for only one of these mutants was studied (dl1457, -12 bp). In this case, the spatial relationship between AAUAAA and poly(A) site is altered. dl1457 produces a class of mRNAs polyadenylated at the first Ca following the AAUAAA sequence, as well as other mRNAs polyadenylated farther downstream. Finally, a 16 bp deletion that includes the AAUAAA sequence prevents poly(A) addition.

Animals↗
PubMed1980

Transcriptional control regions of the adenovirus VAI RNA gene.

By constructing deletion mutations in cloned adenovirus types 2 and 5 VAI genes and measuring the ability of altered templates to direct transcription of VAI RNA in HeLa cell extracts, we have located two transcriptional control regions. The first is an intragenic region located between positions +9 and +72 relative to the 5' end of the VAI(A) RNA. Those deletions examined within these sequences abolished the transcription of mutant templates in HeLa cell extracts. The second control region includes 5' flanking sequences which abut the VAI coding region. Mutations here can reduce the efficiency with which the VAI gene is transcribed. Nucleotide sequence similarities were noted on comparison of the VAI intragenic control region to tRNA sequences, which lead us to speculate that the transcriptional regulation of these two types of genes may be quite similar; the adenovirus VA genes may even have evolved from a tRNA gene(s).

Adenoviruses, Human↗
PubMed1980

Integration in vivo into simian virus 40 DNA of a sequence that resembles a certain family of genomic interspersed repeated sequences.

The isolation and characterization of a viable mutant of simian virus 40 (SV40) called "in1449" are described. the mutant DNA is found to have a 157-nucleotide-long insertion at map position 0.649 within the 5' untranslated sequence of the early region of SV40. The complete nucleotide sequence of the insert is presented. Sequence comparisons show that the insert is not of SV40 origin. The insert is presumably of monkey origin since in1449 was produced within monkey kidney cells. The sequence of the in1449 insert matches remarkably well with sequences of a certain predominant family of interspersed repeated sequences in human DNA (called the Alu family) and cloned members thereof. This high degree of sequence homology suggests that the in1449 insert is derived from a member of a family of interspersed repeated sequences in monkey DNA related to the human Alu family. The in1449 insert (and the Alu family members) contain certain oligonucleotide sequences that also are found conserved in the replication origins of papovaviruses and certain other oligonucleotides found in repetitive double-stranded regions of mammalian heterogeneous nuclear RNAs. Sequences around the two recombinant joints in in1449 exhibit a definite pattern of homology. An octanucleotide present in the SV40 part of the first recombinant joint is exactly repeated 15 nucleotides away within the insert; another octanucleotide present within the insert at the second joint is exactly repeated 21 nucleotides away in the viral DNA. The viral DNA sequences flanking the insert in in1449 also exhibit some homology.

Animals↗
PubMed1979

A mutation which alters initiation of transcription by RNA polymerase III on the Ad5 chromosome.

Mutant dl 309 is a viable Ad5 deletion mutant. Whereas wild-type Ad5-infected HeLa cells contain two VAI RNA species [VAI(A) and VAI(G)] which differ by three nucleotides at their 5' ends, dl 309-infected HeLa cells contain VAI(G) but no VAI(A) RNA. Nucleotide sequence analysis indicates that dl 309 lacks two base pairs which precede the 5' end of VAI(A) by 22 nucleotides. Since the 5' ends of VAI RNAs are not processed, the 309 deletion serves to identify a portion of the sequence required for RNA polymerase III initiation. Since dl 309 grows as well as wild-type Ad5 in HeLa cells, the VAI(A) species is not essential for viral growth in these cells.

Adenoviruses, Human↗