Mutations in the ubiquitin binding UBZ motif of DNA polymerase η do not impair its function in translesion synthesis during replication

Narottam Acharya, Amrita Brahma, Lajos Haracska, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Treatment of Saccharomyces cerevisiae cells with DNA-damaging agents elicits lysine 164-linked PCNA monoubiquitination by Rad6-Rad18. Recently, a number of ubiquitin (Ub) binding domains (UBDs) have been identified in translesion synthesis (TLS) DNA polymerases and it has been proposed that the UBD in a TLS polymerase affects its binding to Ub on PCNA and that this binding mode is indispensable for a TLS polymerase to access PCNA at the site of a stalled replication fork. To evaluate the contribution of the binding of UBDs to the Ub moiety on PCNA in TLS, we have examined the effects of mutations in the C2H2 zinc binding motif and in the conserved D570 residue that lies in the α-helix portion of the UBZ domain of yeast Polη. We find that mutations in the C2H2 motif have no perceptible effect on UV sensitivity or UV mutagenesis, whereas a mutation of the D570 residue adversely affects Polη function. The stimulation of DNA synthesis by Polη with PCNA or Ub-PCNA was not affected by mutations in the C 2H2 motif or the D570 residue. These observations lead us to suggest that the binding of Ub on PCNA via its UBZ domain is not a necessary requirement for the ability of polymerase η to function in TLS during replication.

Original languageEnglish (US)
Pages (from-to)7266-7272
Number of pages7
JournalMolecular and Cellular Biology
Volume27
Issue number20
DOIs
StatePublished - Oct 2007

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Proliferating Cell Nuclear Antigen
DNA-Directed DNA Polymerase
Ubiquitin
Mutation
DNA
Mutagenesis
Lysine
Saccharomyces cerevisiae
Zinc
Yeasts

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Mutations in the ubiquitin binding UBZ motif of DNA polymerase η do not impair its function in translesion synthesis during replication. / Acharya, Narottam; Brahma, Amrita; Haracska, Lajos; Prakash, Louise; Prakash, Satya.

In: Molecular and Cellular Biology, Vol. 27, No. 20, 10.2007, p. 7266-7272.

Research output: Contribution to journalArticle

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