Simian Virus 40 DNA Replication in Vitro

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1984 Nov 1

PMID 6095264

Citations 240

Authors

J J Li

T J Kelly

Affiliations

Soon will be listed here.

Abstract

Soluble extracts prepared from monkey cells (COS-1 or BSC-40) infected with simian virus 40 (SV40) catalyze the efficient replication of exogenously added plasmid DNA molecules containing the cloned SV40 origin of replication. Extracts prepared from uninfected monkey cells also support origin-dependent replication in vitro but only in the presence of added SV40 large tumor (T) antigen. Very little DNA synthesis is observed when the cloned viral origin contains a 4-base-pair deletion mutation known to abolish SV40 DNA replication in vivo or when the parental plasmid vector lacking SV40 sequences is employed as template. The in vitro replication reaction proceeds via branched intermediates (theta structures) that resemble in vivo replication intermediates. Replication is sensitive to aphidicolin but relatively resistant to dideoxythymidine triphosphate. The product of the reaction consists of covalently closed circular DNA molecules that contain full-length daughter strands hydrogen bonded to the parental template. These observations support the conclusion that replication in the in vitro system closely resembles SV40 DNA replication in vivo. The system provides a biochemical assay for the replication activity of SV40 T antigen and should also facilitate the purification and functional characterization of cellular proteins involved in DNA replication.

Citing Articles

Establishing a biochemical understanding of the initiation of chromosome replication in bacteria.

Stillman B Proc Natl Acad Sci U S A. 2024; 121(23):e2400667121.

PMID: 38758693 PMC: 11161774. DOI: 10.1073/pnas.2400667121.

Lagging Strand Initiation Processes in DNA Replication of Eukaryotes-Strings of Highly Coordinated Reactions Governed by Multiprotein Complexes.

Nasheuer H, Onwubiko N Genes (Basel). 2023; 14(5).

PMID: 37239371 PMC: 10218536. DOI: 10.3390/genes14051012.

SV40 T-antigen uses a DNA shearing mechanism to initiate origin unwinding.

Langston L, Yuan Z, Georgescu R, Li H, ODonnell M Proc Natl Acad Sci U S A. 2022; 119(49):e2216240119.

PMID: 36442086 PMC: 9894130. DOI: 10.1073/pnas.2216240119.

Current In Vitro and In Vivo Models to Study MCPyV-Associated MCC.

Loke A, Lambert P, Spurgeon M Viruses. 2022; 14(10).

PMID: 36298759 PMC: 9607385. DOI: 10.3390/v14102204.

Evaluation and modulation of DNA lesion bypass in an SV40 large T antigen-based in vitro replication system.

Szeltner Z, Poti A, Harami G, Kovacs M, Szuts D FEBS Open Bio. 2021; 11(4):1054-1075.

PMID: 33512058 PMC: 8016126. DOI: 10.1002/2211-5463.13099.

References

Shortle D, Margolskee R, Nathans D . Mutational analysis of the simian virus 40 replicon: pseudorevertants of mutants with a defective replication origin. Proc Natl Acad Sci U S A. 1979; 76(12):6128-31. PMC: 411816. DOI: 10.1073/pnas.76.12.6128. View

Fareed G, Garon G, Salzman N . Origin and direction of simian virus 40 deoxyribonucleic acid replication. J Virol. 1972; 10(3):484-91. PMC: 356490. DOI: 10.1128/JVI.10.3.484-491.1972. View

DePamphilis M, Berg P . Requirement of a Cytoplasmic Fraction for Synthesis of SV40 Deoxyribonucleic Acid in Isolated Nuclei*. J Biol Chem. 1975; 250(11):4348-54. View

Lusky M, Botchan M . Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981; 293(5827):79-81. DOI: 10.1038/293079a0. View

Gluzman Y, Frisque R, Sambrook J . Origin-defective mutants of SV40. Cold Spring Harb Symp Quant Biol. 1980; 44 Pt 1,:293-300. DOI: 10.1101/sqb.1980.044.01.033. View

DePamphilis M, Beard P, Berg P . Synthesis of Superhelical Simian Virus 40 Deoxyribonucleic Acid in Cell Lysates*. J Biol Chem. 1975; 250(11):4340-7. View

Challberg M, Kelly Jr T . Adenovirus DNA replication in vitro. Proc Natl Acad Sci U S A. 1979; 76(2):655-9. PMC: 383008. DOI: 10.1073/pnas.76.2.655. View

Edenberg H, Anderson S, DePamphilis M . Involvement of DNA polymerase alpha in simian virus 40 DNA replication. J Biol Chem. 1978; 253(9):3273-80. View

Griffith J . Chromatin structure: deduced from a minichromosome. Science. 1975; 187(4182):1202-3. DOI: 10.1126/science.187.4182.1202. View

10.

Challberg M, Kelly T . Eukaryotic DNA replication: viral and plasmid model systems. Annu Rev Biochem. 1982; 51:901-34. DOI: 10.1146/annurev.bi.51.070182.004345. View

11.

Danna K, Nathans D . Bidirectional replication of Simian Virus 40 DNA. Proc Natl Acad Sci U S A. 1972; 69(11):3097-100. PMC: 389712. DOI: 10.1073/pnas.69.11.3097. View

12.

Cremisi C, Pignatti P, CROISSANT O, Yaniv M . Chromatin-like structures in polyoma virus and simian virus 10 lytic cycle. J Virol. 1975; 17(1):204-11. PMC: 515404. DOI: 10.1128/JVI.17.1.204-211.1976. View

13.

Myers R, Tjian R . Construction and analysis of simian virus 40 origins defective in tumor antigen binding and DNA replication. Proc Natl Acad Sci U S A. 1980; 77(11):6491-5. PMC: 350311. DOI: 10.1073/pnas.77.11.6491. View

14.

Challberg M, Kelly Jr T . Adenovirus DNA replication in vitro: origin and direction of daughter strand synthesis. J Mol Biol. 1979; 135(4):999-1012. DOI: 10.1016/0022-2836(79)90524-2. View

15.

DiMaio D, Nathans D . Regulatory mutants of simian virus 40. Effect of mutations at a T antigen binding site on DNA replication and expression of viral genes. J Mol Biol. 1982; 156(3):531-48. DOI: 10.1016/0022-2836(82)90265-0. View

16.

Edenberg H, Waqar M, Huberman J . DNA synthesis by partially purified replicating simian virus 40 chromosomes. Nucleic Acids Res. 1977; 4(9):3083-95. PMC: 342636. DOI: 10.1093/nar/4.9.3083. View

17.

SEBRING E, Kelly Jr T, THOREN M, Salzman N . Structure of replicating simian virus 40 deoxyribonucleic acid molecules. J Virol. 1971; 8(4):478-90. PMC: 376221. DOI: 10.1128/JVI.8.4.478-490.1971. View

18.

Peden K, Pipas J, Pearson-White S, Nathans D . Isolation of mutants of an animal virus in bacteria. Science. 1980; 209(4463):1392-6. DOI: 10.1126/science.6251547. View

19.

Su R, DePamphilis M . In vitro replication of simian virus 40 DNA in a nucleoprotein complex. Proc Natl Acad Sci U S A. 1976; 73(10):3466-70. PMC: 431136. DOI: 10.1073/pnas.73.10.3466. View

20.

Ariga H, Sugano S . Initiation of simian virus 40 DNA replication in vitro. J Virol. 1983; 48(2):481-91. PMC: 255373. DOI: 10.1128/JVI.48.2.481-491.1983. View