Interaction of hREV1 with three human Y-family DNA polymerases

Genes Cells. 2004 Jun;9(6):523-31. doi: 10.1111/j.1356-9597.2004.00747.x.

Abstract

Polkappa is one of many DNA polymerases involved in translesion DNA synthesis (TLS). It belongs to the Y-family of polymerases along with Poleta, Poliota and hREV1. Unlike Poleta encoded by the xeroderma pigmentosum variant (XPV) gene, Polkappa is unable to bypass UV-induced DNA damage in vitro, but it is able to bypass benzo[a]pyrene (B[a]P)-adducted guanines accurately and efficiently. In an attempt to identify factor(s) targeting Polkappa to its cognate DNA lesion(s), we searched for Polkappa-interacting proteins by using the yeast two-hybrid assay. We found that Polkappa interacts with a C-terminal region of hREV1. Poleta and Poliota were also found to interact with the same region of hREV1. The interaction between Polkappa and hREV1 was confirmed by pull-down and co-immunoprecipitation assays. The C-terminal region of hREV1 is known to interact with hREV7, a non-catalytic subunit of Polzeta that is another structurally unrelated TLS enzyme, and we show that Polkappa and hREV7 bind to the same C-terminal region of hREV1. Thus, our results suggest that hREV1 plays a pivotal role in the multi-enzyme, multi-step process of translesion DNA synthesis.

MeSH terms

  • Amino Acid Sequence
  • Benzo(a)pyrene / pharmacology
  • Cell Line
  • DNA Damage / drug effects
  • DNA Damage / genetics*
  • DNA Damage / radiation effects
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / classification
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Escherichia coli / genetics
  • Fibroblasts / metabolism
  • Glutathione Transferase / metabolism
  • Guanine / metabolism
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins
  • Nucleotidyltransferases / chemistry
  • Nucleotidyltransferases / genetics*
  • Nucleotidyltransferases / metabolism*
  • Precipitin Tests
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion
  • Two-Hybrid System Techniques
  • Ultraviolet Rays
  • beta-Galactosidase / metabolism

Substances

  • Nuclear Proteins
  • Recombinant Fusion Proteins
  • Benzo(a)pyrene
  • Guanine
  • Glutathione Transferase
  • DNA polymerase iota
  • Nucleotidyltransferases
  • REV1 protein, human
  • DNA-Directed DNA Polymerase
  • POLK protein, human
  • beta-Galactosidase