Electron microscopic analysis reveals that replication factor C is sequestered by single-stranded DNA

Nucleic Acids Res. 1999 Sep 1;27(17):3433-7. doi: 10.1093/nar/27.17.3433.

Abstract

Replication factor C (RF-C) is a eukaryotic heteropentameric protein required for DNA replication and repair processes by loading proliferating cell nuclear antigen (PCNA) onto DNA in an ATP-dependent manner. Prior to loading PCNA, RF-C binds to DNA. This binding is thought to be restricted to a specific DNA structure, namely to a primer/template junction. Using the electron microscope we have examined the affinity of human heteropentameric RF-C and the DNA-binding region within the large subunit of RF-C from Drosophila melanogaster (dRF-Cp140) to heteroduplex DNA. The electron microscopic data indicate that both human heteropentameric RF-C and the DNA-binding region within dRF-Cp140 are sequestered by single-stranded DNA. No preferential affinity for the 3' or 5' transition points from single- to double-stranded DNA was evident.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Single-Stranded / chemistry
  • DNA, Single-Stranded / physiology*
  • DNA, Single-Stranded / ultrastructure
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / physiology*
  • DNA-Binding Proteins / ultrastructure
  • Drosophila / chemistry
  • HeLa Cells
  • Homeodomain Proteins*
  • Humans
  • Microscopy, Electron*
  • Minor Histocompatibility Antigens
  • Nucleic Acid Heteroduplexes / ultrastructure
  • Plasmids / chemistry
  • Protein Binding
  • Proto-Oncogene Proteins c-bcl-2*
  • Replication Protein C
  • Repressor Proteins*
  • Saccharomyces cerevisiae Proteins*

Substances

  • BCL2-related protein A1
  • DNA, Single-Stranded
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • MATA1 protein, S cerevisiae
  • Minor Histocompatibility Antigens
  • Nucleic Acid Heteroduplexes
  • Proto-Oncogene Proteins c-bcl-2
  • Repressor Proteins
  • Saccharomyces cerevisiae Proteins
  • Replication Protein C