Interaction of AD2+D2 protein and simian virus 40 large T antigen with the large tumor antigen binding site I

Biochemistry ◽  
1984 ◽  
Vol 23 (25) ◽  
pp. 5938-5944 ◽  
Author(s):  
Eric F. Fisher ◽  
Patricia L. Feist ◽  
Serge L. Beaucage ◽  
Richard M. Meyers ◽  
Robert Tjian ◽  
...  
Keyword(s):  
Binding Site ◽  
Tumor Antigen ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Antigen Binding ◽  
Large T Antigen ◽  
Antigen Binding Site ◽  
Large Tumor ◽  
D2 Protein

scholarly journals New Insights into the Mechanism of Inhibition of p53 by Simian Virus 40 Large T Antigen

1999 ◽  
Vol 19 (4) ◽  
pp. 2746-2753 ◽  
Author(s):  
Hilary M. Sheppard ◽  
Siska I. Corneillie ◽  
Christine Espiritu ◽  
Andrea Gatti ◽  
Xuan Liu
Keyword(s):  
Tumor Antigen ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Human Cells ◽  
Transactivation Domain ◽  
Large T Antigen ◽  
Large Tumor ◽  
Mechanism Of Inhibition ◽  
Mouse Cells

ABSTRACT Simian virus 40 (SV40) large tumor antigen (T antigen) has been shown to inhibit p53-dependent transcription by preventing p53 from binding to its cognate cis element. Data presented in this report provide the first direct functional evidence that T antigen, under certain conditions, may also repress p53-dependent transcription by a mechanism in which the transactivation domain of p53 is abrogated while DNA binding is unaffected. Specifically, p53 purified as a complex with T antigen from mouse cells was found to bind DNA as a transcriptionally inactive intact complex, while that purified from human cells was found to bind DNA independently of T antigen and could activate p53-dependent transcription. This difference in activity may be dependent on a different interaction of T antigen with mouse and human p53 and, in addition, on the presence of super T, which is found only in transformed rodent cells. These results suggest that subtle yet important differences exist between the inhibition of p53 by T antigen in mouse and human cells. The implications of this finding with respect to SV40-associated malignancies are discussed.

Sequence-specific interactions between a cellular DNA-binding protein and the simian virus 40 origin of DNA replication

1988 ◽  
Vol 8 (2) ◽  
pp. 903-911
Author(s):  
W Traut ◽  
E Fanning
Keyword(s):  
Dna Replication ◽  
Binding Site ◽  
Base Pair ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Antigen Binding ◽  
Antigen Binding Site ◽  
Pair Sequence ◽  
Sv40 Dna

The core origin of simian virus 40 (SV40) DNA replication is composed of a 64-base-pair sequence encompassing T-antigen-binding site II and adjacent sequences on either side. A 7-base-pair sequence to the early side of T-antigen-binding site II which is conserved among the papovavirus genomes SV40, BK, JC, and SA12 was recently shown to be part of a 10-base-pair sequence required for origin activity (S. Deb, A.L. DeLucia, C.-P. Baur, A. Koff, and P. Tegtmeyer, Mol. Cell. Biol. 6:1663-1670, 1986), but its functional role was not defined. In the present report, we have used gel retention assays to identify a monkey cell factor that interacts specifically with double-stranded DNA carrying this sequence and also binds to single-stranded DNA. DNA-protein complexes formed with extracts from primate cells are more abundant and display electrophoretic mobilities distinct from those formed with rodent cell extracts. The binding activity of the factor on mutant templates is correlated with the replication activity of the origin. The results suggest that the monkey cell factor may be involved in SV40 DNA replication.

Homologous recombination enhancement conferred by the Z-DNA motif d(TG)30 is abrogated by simian virus 40 T antigen binding to adjacent DNA sequences

1990 ◽  
Vol 10 (2) ◽  
pp. 794-800
Author(s):  
W P Wahls ◽  
P D Moore
Keyword(s):  
Homologous Recombination ◽  
Binding Site ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Antigen Binding ◽  
Antigen Binding Site ◽  
Left Handed ◽  
Dna Motif ◽  
Z Dna

The Z-DNA motif polydeoxythymidylic-guanylic [d(TG)].polydeoxyadenylic-cytidylic acid [d(AC)], present throughout eucaryotic genomes, is capable of readily forming left-handed Z-DNA in vitro and has been shown to promote homologous recombination. The effects of simian virus 40 T-antigen-dependent substrate replication upon the stimulation of recombination conferred by the Z-DNA motif d(TG)30 were analyzed. Presence of d(TG)30 adjacent to a T-antigen-binding site I can stimulate homologous recombination between nonreplicating plasmids, providing that T antigen is absent, in both simian CV-1 cells and human EJ cells (W. P. Wahls, L. J. Wallace, and P. D. Moore, Mol. Cell. Biol. 10:785-793). It has also been shown elsewhere that the presence of d(TG)n not adjacent to the T-antigen-binding site can stimulate homologous recombination in simian virus 40 molecules replicating in the presence of T antigen (P. Bullock, J. Miller, and M. Botchan, Mol. Cell. Biol. 6:3948-3953, 1986). However, it is demonstrated here that d(TG)30 nine base pairs distant from a T-antigen-binding site bound with T antigen does not stimulate recombination between either replicating or nonreplicating substrates in somatic cells. The bound T antigen either prevents the d(TG)30 sequence from acquiring a recombinogenic configuration (such as left-handed Z-DNA), or it prevents the interaction of recombinase proteins with the sequence by stearic hindrance.

scholarly journals Homologous recombination enhancement conferred by the Z-DNA motif d(TG)30 is abrogated by simian virus 40 T antigen binding to adjacent DNA sequences.

1990 ◽  
Vol 10 (2) ◽  
pp. 794-800 ◽  
Author(s):  
W P Wahls ◽  
P D Moore
Keyword(s):  
Homologous Recombination ◽  
Binding Site ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Antigen Binding ◽  
Antigen Binding Site ◽  
Left Handed ◽  
Dna Motif ◽  
Z Dna

The Z-DNA motif polydeoxythymidylic-guanylic [d(TG)].polydeoxyadenylic-cytidylic acid [d(AC)], present throughout eucaryotic genomes, is capable of readily forming left-handed Z-DNA in vitro and has been shown to promote homologous recombination. The effects of simian virus 40 T-antigen-dependent substrate replication upon the stimulation of recombination conferred by the Z-DNA motif d(TG)30 were analyzed. Presence of d(TG)30 adjacent to a T-antigen-binding site I can stimulate homologous recombination between nonreplicating plasmids, providing that T antigen is absent, in both simian CV-1 cells and human EJ cells (W. P. Wahls, L. J. Wallace, and P. D. Moore, Mol. Cell. Biol. 10:785-793). It has also been shown elsewhere that the presence of d(TG)n not adjacent to the T-antigen-binding site can stimulate homologous recombination in simian virus 40 molecules replicating in the presence of T antigen (P. Bullock, J. Miller, and M. Botchan, Mol. Cell. Biol. 6:3948-3953, 1986). However, it is demonstrated here that d(TG)30 nine base pairs distant from a T-antigen-binding site bound with T antigen does not stimulate recombination between either replicating or nonreplicating substrates in somatic cells. The bound T antigen either prevents the d(TG)30 sequence from acquiring a recombinogenic configuration (such as left-handed Z-DNA), or it prevents the interaction of recombinase proteins with the sequence by stearic hindrance.

scholarly journals Sequence-specific interactions between a cellular DNA-binding protein and the simian virus 40 origin of DNA replication.

1988 ◽  
Vol 8 (2) ◽  
pp. 903-911 ◽  
Author(s):  
W Traut ◽  
E Fanning
Keyword(s):  
Dna Replication ◽  
Binding Site ◽  
Base Pair ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Antigen Binding ◽  
Antigen Binding Site ◽  
Pair Sequence ◽  
Sv40 Dna

The core origin of simian virus 40 (SV40) DNA replication is composed of a 64-base-pair sequence encompassing T-antigen-binding site II and adjacent sequences on either side. A 7-base-pair sequence to the early side of T-antigen-binding site II which is conserved among the papovavirus genomes SV40, BK, JC, and SA12 was recently shown to be part of a 10-base-pair sequence required for origin activity (S. Deb, A.L. DeLucia, C.-P. Baur, A. Koff, and P. Tegtmeyer, Mol. Cell. Biol. 6:1663-1670, 1986), but its functional role was not defined. In the present report, we have used gel retention assays to identify a monkey cell factor that interacts specifically with double-stranded DNA carrying this sequence and also binds to single-stranded DNA. DNA-protein complexes formed with extracts from primate cells are more abundant and display electrophoretic mobilities distinct from those formed with rodent cell extracts. The binding activity of the factor on mutant templates is correlated with the replication activity of the origin. The results suggest that the monkey cell factor may be involved in SV40 DNA replication.

scholarly journals Replication but Not Transcription of Simian Virus 40 DNA Is Dependent on Nuclear Domain 10

2000 ◽  
Vol 74 (20) ◽  
pp. 9694-9700 ◽  
Author(s):  
Qiyi Tang ◽  
Peter Bell ◽  
Peter Tegtmeyer ◽  
Gerd G. Maul
Keyword(s):  
Simian Virus 40 ◽  
Adenovirus Type ◽  
Simian Virus ◽  
T Antigen ◽  
Large T Antigen ◽  
Antigen Binding Site ◽  
Viral Origin ◽  
Coding Sequence ◽  
Nuclear Domain ◽  
Transcription And Replication

ABSTRACT DNA viruses from several families including herpes simplex virus type 1, adenovirus type 5, and simian virus 40 (SV40), start their transcription and replication adjacent to a specific nuclear domain, ND10. We asked whether a specific viral DNA sequence determines the location of these synthetic activities at such restricted nuclear sites. Partial and overlapping SV40 sequences were introduced into a β-galactosidase expression vector, and the β-galactosidase transcripts were localized by in situ hybridization. Transcripts derived from control plasmids were found throughout the nucleus and at highly concentrated sites but not at ND10. SV40 genomic segments supported ND10-associated transcription only when the origin and the coding sequence for the large T antigen were present. When the large T-antigen coding sequence was eliminated but the T antigen was constitutively expressed in COS-7 cells, the viral origin was sufficient to localize transcription and replication to ND10. Deletion analysis showed that only the large T-antigen binding site II (the core origin) was required but the T antigen was needed for detectable transcription at ND10. Large T antigen expressed from plasmids without the viral core origin did not bind or localize to ND10. Blocking of DNA replication prevented the accumulation of transcripts at ND10, indicating that only sites with replicating templates accumulated transcripts. Transcription at ND10 did not enhance total protein synthesis of plasmid transcripts. These findings suggest that viral transcription at ND10 may only be a consequence of viral genomes directed to ND10 for replication. Although plasmid transcription can take place anywhere in the nucleus, T-antigen-directed replication is apparently restricted to ND10.

trans Activation of the simian virus 40 late transcription unit by T-antigen

1985 ◽  
Vol 5 (6) ◽  
pp. 1391-1399
Author(s):  
J Brady ◽  
G Khoury
Keyword(s):  
Gene Expression ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Transcription Unit ◽  
T Antigen ◽  
Late Gene ◽  
Antigen Binding ◽  
Late Gene Expression ◽  
Antigen Binding Site

We have investigated the role of simian virus 40 (SV40) T-antigen in the induction of late gene expression independent of its function in amplifying templates through DNA replication. Northern blot and S1 nuclease analyses showed that stimulation occurred at the transcriptional level. At least two template elements, the T-antigen-binding sites and the 72-base-pair repeats, appeared to be important for this induction. Using template mutants, we demonstrated that deletions within T-antigen-binding site II decreased T-antigen-mediated late gene expression approximately 10- to 20-fold. In addition, multiple point mutations within a single retained copy of the SV40 72-base-pair repeat decreased T-antigen-mediated late gene expression. Using in vivo competition studies, we demonstrated that competitor DNA fragments containing the SV40 control region (nucleotides 5171 through 272) quantitatively decreased SV40 late gene expression in COS-1 cells. In contrast, competition with a plasmid containing SV40 nucleotides 1 through 294 (which removes all of T-antigen-binding site I and half of site II) was much less efficient. Finally, we demonstrated that in vivo competition experiments employing competitor fragments distal to the T-antigen-binding sites within the late template region (SV40 nucleotides 180 through 2533) resulted in superinduction of late gene expression in COS-1 cells. This finding suggests that negative factors such as repressors or attenuators may modulate late SV40 gene expression before induction. Our results are consistent with a model in which induction of late gene expression involves an interaction of the SV40 origin region with DNA-binding proteins, one of which may be T-antigen. Activation of the SV40 late transcription unit may involve induction of the SV40 enhancer or removal of a repressor-like protein or both.

scholarly journals Stabilization of the 53,000-dalton nonviral tumor antigen is not required for transformation by simian virus 40.

1983 ◽  
Vol 3 (2) ◽  
pp. 290-296 ◽  
Author(s):  
L M Sompayrac ◽  
E G Gurney ◽  
K J Danna
Keyword(s):  
Cell Lines ◽  
Tumor Antigen ◽  
Deletion Mutant ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Large T Antigen ◽  
Coding Region ◽  
Present Evidence ◽  
Mouse Cells

We have isolated a simian virus 40 deletion mutant, F8dl, that lacks the sequences from 0.168 to 0.424 map units. The deleted sequences represent about one-half of the coding region for large T antigen. We present evidence here that F8dl is able to transform mouse cells in a focus assay and that cell lines derived from these foci exhibit fully transformed phenotypes, have integrated mutant genomes, and express mutant-encoded proteins. This result implies that the region of the simian virus 40 genome between 0.168 and 0.424 map units is not essential for the maintenance of transformation. In addition, we have found that cells fully transformed by F8dl produce a 53,000-dalton nonviral tumor antigen (p53) that is as unstable as the p53 of untransformed cells. From this result we infer that transformation by simian virus 40 does not require the stabilization of p53.

scholarly journals Production of simian virus 40 large tumor antigen in bacteria: altered DNA-binding specificity and dna-replication activity of underphosphorylated large tumor antigen

1989 ◽  
Vol 86 (17) ◽  
pp. 6479-6483 ◽  
Author(s):  
I J Mohr ◽  
Y Gluzman ◽  
M P Fairman ◽  
M Strauss ◽  
D McVey ◽  
...  
Keyword(s):  
Tumor Antigen ◽  
Mammalian Cells ◽  
Expression System ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Origin Of Replication ◽  
Wild Type ◽  
Large Tumor ◽  
Large Tumor Antigen

A bacterial expression system was used to produce simian virus 40 large tumor antigen (T antigen) in the absence of the extensive posttranslational modifications that occur in mammalian cells. Wild-type T antigen produced in bacteria retained a specific subset of the biochemical activities displayed by its mammalian counterpart. Escherichia coli T antigen functioned as a helicase and bound to DNA fragments containing either site I or the wild-type origin of replication in a manner identical to mammalian T antigen. However, T antigen purified from E. coli did not efficiently bind to site II, an essential cis element within the simian virus 40 origin of replication. It therefore could not unwind origin-containing plasmids or efficiently replicate simian virus 40 DNA in vitro. The ability of protein phosphorylation to modulate the intrinsic preference of full-length T antigen for either site I or site II is discussed.

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