Sequence and expression in Escherichia coli of the 40-kDa subunit of activator 1 (replication factor C) of HeLa cells

Mei Chen, Z. Q. Pan, Jerard Hurwitz

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

Activator 1 (A1; also called replication factor C), in conjunction with proliferating-cell nuclear antigen (PCNA), is essential for the elongation of primed DNA templates by DNA polymerases δ and ε. A1 contains five distinct subunits of 145, 40, 38, 37, and 36.5 kDa. Here we describe the isolation, sequence, and bacterial expression of a cDNA coding for the 40-kDa subunit. In keeping with the presence of an ATP-binding motif, the bacterially expressed 40-kDa subunit binds ATP. The interaction between the 40-kDa subunit and ATP was reduced by the addition of PCNA. In addition, antibodies raised against the 40-kDa subunit abolished the A1-and PCNA-dependent synthesis of DNA catalyzed by polymerase δ. The putative amino acid sequence of the 40-kDa subunit of A1 revealed significant homology with the bacteriophage T4 gene 44 protein and, to a lesser degree, with the τ and γ subunits of Escherichia coli DNA polymerase III holoenzyme.

Original languageEnglish
Pages (from-to)2516-2520
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number7
DOIs
StatePublished - 1992
Externally publishedYes

Keywords

  • DNA polymerases δ and ε
  • DNA synthesis
  • Proliferating-cell nuclear antigen

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