Identification of cellular proteins required for simian virus 40 DNA replication.

Published

Journal Article

Study of the proteins involved in DNA replication of a model system such as SV40 is a first step in understanding eukaryotic chromosomal replication. Using a cell-free system that is capable of replicating plasmid DNA molecules containing the SV40 origin of replication, we conducted a series of systematic fractionation-reconstitution experiments for the purpose of identifying and characterizing the cellular proteins involved in SV40 DNA replication. In addition to the one viral-encoded replication protein, T antigen, we have identified and begun to characterize at least six cellular components from a HeLa cytoplasmic extract that are absolutely required for SV40 DNA replication in vitro. These include: (i) two partially purified fractions, CF IC and CF IIA, and (ii) four proteins that have been purified to near homogeneity, replication protein-A, proliferating cell nuclear antigen, DNA polymerase alpha-primase complex, and topoisomerase (I and II). Replication protein-A is a multi-subunit protein that has single-stranded DNA binding activity and is required for a T antigen-dependent, origin-dependent unwinding reaction which may be an important early step in initiation of replication. Fraction CF IC can stimulate this unwinding reaction, suggesting that it also may function during initiation. Proliferating cell nuclear antigen, DNA polymerase alpha-primase, and CF IIA all appear to be involved in elongation of nascent chains.

Full Text

Duke Authors

Cited Authors

  • Wold, MS; Weinberg, DH; Virshup, DM; Li, JJ; Kelly, TJ

Published Date

  • February 15, 1989

Published In

Volume / Issue

  • 264 / 5

Start / End Page

  • 2801 - 2809

PubMed ID

  • 2536723

Pubmed Central ID

  • 2536723

International Standard Serial Number (ISSN)

  • 0021-9258

Language

  • eng

Conference Location

  • United States