Binding and unwinding—How T antigen engages the SV40 origin of DNA replication

Borowiec, James A.; Dean, Frank B.; Bullock, Peter A.; Hurwitz, Jerard

doi:10.1016/0092-8674(90)90730-3

Cited by 386 publications

(282 citation statements)

References 21 publications

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“…7B shows the maximal stimulation achieved with each pair of reciprocal exchange mutants. Firstly, we observed that a complex containing a murine p180 subunit, MH 3 , fails almost entirely to be stimulated by SV40 TAg. Secondly, we observed that there exists among all complexes a qualitative correlation between those that show a strong TAg-mediated stimulation in this assay and those that function well in the FIG.…”

Section: H671m Complexes (Lanesmentioning

confidence: 95%

“…Polyomavirus DNA replication has been found to be largely dependent upon factors involved in replication of the host DNA but does contribute one essential trans-acting factor known as large T antigen (TAg) to the replication complex. TAg is responsible for recognition of the viral origin of replication, at which it forms a double hexamer (3). In the presence of the single-stranded DNA-binding protein, RPA (replication protein A), and topoisomerase I, TAg proceeds to unwind the origin DNA and recruits the DNA polymerase ␣-primase heterotetramer, which then initiates bidirectional DNA synthesis (4 -11).…”

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confidence: 99%

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Species Specificity of Simian Virus 40 DNA Replication in Vitro Requires Multiple Functions of Human DNA Polymerase α

Smith¹,

Steffen²,

Grosse³

et al. 2002

Journal of Biological Chemistry

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Human cell extracts support the replication of SV40 DNA, whereas mouse cell extracts do not. Species specificity is determined at the level of initiation of DNA replication, and it was previously found that this requires the large subunit, p180, of DNA polymerase ␣-primase to be of human origin. Furthermore, a functional interaction between SV40 large T antigen (TAg) and p180 is essential for viral DNA replication. In this study we determined that the N-terminal regions of human p180, which contain the TAg-binding sites, can be replaced with those of murine origin without losing the ability to support SV40 DNA replication in vitro. The same substitutions do not prevent SV40 TAg from stimulating the activity of DNA polymerase ␣-primase on single-stranded DNA in the presence of replication protein A. Furthermore, biophysical studies show that the interactions of human and murine DNA polymerase ␣-primase with SV40 TAg are of a similar magnitude. These studies strongly suggest that requirement of SV40 DNA replication for human DNA polymerase ␣ depends neither on the TAg-binding site being of human origin nor on the strength of the binary interaction between SV40 TAg and DNA polymerase ␣-primase but rather on sequences in the C-terminal region of human p180.Polyomavirus DNA replication has been studied extensively because of the ease with which viruses of this family, which include SV40 and mouse polyoma virus (PyV), 1 can be grown in cell culture and moreover because of the availability of an in vitro replication system, which allows detailed investigation of the individual factors that play a role in the replication process (1, 2). Polyomavirus DNA replication has been found to be largely dependent upon factors involved in replication of the host DNA but does contribute one essential trans-acting factor known as large T antigen (TAg) to the replication complex. TAg is responsible for recognition of the viral origin of replication, at which it forms a double hexamer (3). In the presence of the single-stranded DNA-binding protein, RPA (replication protein A), and topoisomerase I, TAg proceeds to unwind the origin DNA and recruits the DNA polymerase ␣-primase heterotetramer, which then initiates bidirectional DNA synthesis (4 -11). DNA polymerase ␣-primase initiates DNA replication through the action of its smallest subunit, p48, which synthesizes RNA primers that are subsequently elongated by the large subunit, p180 (12-15). The bulk of DNA synthesis is, however, carried out by a more processive enzyme complex consisting of DNA polymerase ␦ and its processivity factor, PCNA (16 -18). The transition between DNA synthesis by DNA polymerases ␣ and ␦ is mediated by the PCNA loading factor replication factor C (16, 19). Throughout the viral replication cycle, TAg double hexamers are thought to act as the replicative helicase (20). For a more extensive account of the DNA replication process and its participating factors see Refs. 1, 2, and 21-23. SV40 and PyV are very similar with regard to DNA replication; their respectiv...

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Section: H671m Complexes (Lanesmentioning

confidence: 95%

mentioning

confidence: 99%

Species Specificity of Simian Virus 40 DNA Replication in Vitro Requires Multiple Functions of Human DNA Polymerase α

Smith¹,

Steffen²,

Grosse³

et al. 2002

Journal of Biological Chemistry

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“…Precipitations were carried out either directly after tryptic cleavage (lanes 2, 3, 6, 7, 10, 11, 14, and 15) or after denaturing the sample (lanes 4, 8, 12, and 16). The products were separated by 13.5% SDS-PAGE and analyzed by immunoblotting with Pab204 (lanes [1][2][3][4][5][6][7][8] or Pab419 (lanes 9 -16). Peptide numbers are indicated on the left, and those that co-precipitated after cleavage, but not after denaturation (den.…”

Section: Limited Proteolysis Of Tag Hexamers and Doublementioning

confidence: 99%

Partial Proteolysis of Simian Virus 40 T Antigen Reveals Intramolecular Contacts between Domains and Conformation Changes upon Hexamer Assembly

Weißhart¹,

Friedl²,

Taneja

et al. 2004

Journal of Biological Chemistry

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“…In the first steps of DNA replication, specific proteins are involved in origin recognition and unwinding of the DNA. Additional proteins such as replication factor A (RF-A) are needed to stabilize this complex [4]. The initiation of DNA synthesis requires the interaction of DNA polymerase c1 with the RF-A complex.…”

Section: Introductionmentioning

confidence: 99%

Nuclear recruitment of A l p 145 subunit of replication factor C in the early GI phase of the cell cycle in Faza 567 hepatoma cell line and hepatocyte primary cultures

et al. 1995

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Using a combination of immunoprecipitation and Western blotting with Faza 567 hepatoma cell extracts revealed that the large subunit of replication factor C (Alp145; mRFC140) was in a complex with proliferating cell nuclear antigen (PCNA). Western blotting showed that Alp145 was more abundant in nuclear extracts from butyrate-treated hepatoma cells which blocks the cells in the Cl phase of the cell cycle than from routinely cultured cells. Indirect immunoperoxidase analysis of Cl blocked Faza hepatoma cells localized Alp145 protein predominantly in the nucleoli. When hepatoma cells were stimulated to progress toward the S phase, Alp145 protein was then observed in both the cytoplasm and the nucleoplasm of these cells. Studies with early cultured normal hepatocytes which are progressing from Go towards G,, also showed a nucleolus distribution for Alp145. This is the first demonstration in mammalian cells that the large subunit of replication factor C is associated with PCNA in the nucleus and that its distribution within cells changes during the cell cycle.

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Binding and unwinding—How T antigen engages the SV40 origin of DNA replication

Cited by 386 publications

References 21 publications

Species Specificity of Simian Virus 40 DNA Replication in Vitro Requires Multiple Functions of Human DNA Polymerase α

Species Specificity of Simian Virus 40 DNA Replication in Vitro Requires Multiple Functions of Human DNA Polymerase α

Partial Proteolysis of Simian Virus 40 T Antigen Reveals Intramolecular Contacts between Domains and Conformation Changes upon Hexamer Assembly

Nuclear recruitment of A l p 145 subunit of replication factor C in the early GI phase of the cell cycle in Faza 567 hepatoma cell line and hepatocyte primary cultures

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