The C-terminal zinc finger of the catalytic subunit of DNA polymerase delta is responsible for direct interaction with the B-subunit

JS Garcia, LF Ciufo, XW Yang, SE Kearsey, Stuart Andrew MacNeill

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55 Citations (Scopus)

Abstract

DNA polymerase delta (Pol delta) plays a central role in eukaryotic chromosomal DNA replication, repair and recombination. In fission yeast, Pol delta is a tetrameric enzyme, comprising the catalytic subunit Pol3 and three smaller subunits, Cdc1, Cdc27 and Cdm1. Previous studies have demonstrated a direct interaction between Pol3 and Cdc1, the B-subunit of the complex. Here it is shown that removal of the tandem zinc finger modules located at the C-terminus of Pol3 by targeted proteolysis renders the Pol3 protein non-functional in vivo, and that the C-terminal zinc finger module ZnF2 is both necessary and sufficient for binding to the B-subunit in vivo and in vitro. Extensive mutagenesis of the ZnF2 module identifies important residues for B-subunit binding. In particular, disruption of the ZnF2 module by substitution of the putative metal-coordinating cysteines with alanine abolishes B-subunit binding and in vivo function. Finally, evidence is presented suggesting that the ZnF region is post-translationally modified in fission yeast cells.

Original languageEnglish
Pages (from-to)3005-3016
Number of pages12
JournalNucleic Acids Research
Volume32
Issue number10
DOIs
Publication statusPublished - Jun 2004

Keywords

  • SCHIZOSACCHAROMYCES-POMBE
  • SACCHAROMYCES-CEREVISIAE
  • FISSION YEAST
  • REPLICATION FORK
  • RECONSTITUTION
  • EPSILON
  • HOMOLOG
  • GENE
  • MUTAGENESIS
  • DIVISION

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