Mutations at the Subunit Interface of Yeast Proliferating Cell Nuclear Antigen Reveal a Versatile Regulatory Domain

Miklos Halmai, Orsolya Frittmann, Zoltan Szabo, Andreea Daraba, Vamsi K. Gali, Eva Balint, Ildiko Unk

Research output: Contribution to journalArticle

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Abstract

Proliferating cell nuclear antigen (PCNA) plays a key role in many cellular processes and due to that it interacts with a plethora of proteins. The main interacting surfaces of Saccharomyces cerevisiae PCNA have been mapped to the interdomain connecting loop and to the carboxy-terminal domain. Here we report that the subunit interface of yeast PCNA also has regulatory roles in the function of several DNA damage response pathways. Using site-directed mutagenesis we engineered mutations at both sides of the interface and investigated the effect of these alleles on DNA damage response. Genetic experiments with strains bearing the mutant alleles revealed that mutagenic translesion synthesis, nucleotide excision repair, and homologous recombination are all regulated through residues at the subunit interface. Moreover, genetic characterization of one of our mutants identifies a new sub-branch of nucleotide excision repair. Based on these results we conclude that residues at the subunit boundary of PCNA are not only important for the formation of the trimer structure of PCNA, but they constitute a regulatory protein domain that mediates different DNA damage response pathways, as well.

Original languageEnglish
Article numbere0161307
Pages (from-to)1-20
Number of pages20
JournalPloS ONE
Volume11
Issue number8
DOIs
Publication statusPublished - 18 Aug 2016

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proliferating cell nuclear antigen
Proliferating Cell Nuclear Antigen
Yeast
Yeasts
yeasts
mutation
Mutation
DNA damage
DNA Damage
DNA repair
DNA Repair
DNA
Repair
Bearings (structural)
Nucleotides
Alleles
alleles
mutants
Mutagenesis
Homologous Recombination

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Mutations at the Subunit Interface of Yeast Proliferating Cell Nuclear Antigen Reveal a Versatile Regulatory Domain. / Halmai, Miklos; Frittmann, Orsolya; Szabo, Zoltan; Daraba, Andreea; Gali, Vamsi K.; Balint, Eva; Unk, Ildiko.

In: PloS ONE, Vol. 11, No. 8, e0161307, 18.08.2016, p. 1-20.

Research output: Contribution to journalArticle

Halmai, Miklos ; Frittmann, Orsolya ; Szabo, Zoltan ; Daraba, Andreea ; Gali, Vamsi K. ; Balint, Eva ; Unk, Ildiko. / Mutations at the Subunit Interface of Yeast Proliferating Cell Nuclear Antigen Reveal a Versatile Regulatory Domain. In: PloS ONE. 2016 ; Vol. 11, No. 8. pp. 1-20.
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abstract = "Proliferating cell nuclear antigen (PCNA) plays a key role in many cellular processes and due to that it interacts with a plethora of proteins. The main interacting surfaces of Saccharomyces cerevisiae PCNA have been mapped to the interdomain connecting loop and to the carboxy-terminal domain. Here we report that the subunit interface of yeast PCNA also has regulatory roles in the function of several DNA damage response pathways. Using site-directed mutagenesis we engineered mutations at both sides of the interface and investigated the effect of these alleles on DNA damage response. Genetic experiments with strains bearing the mutant alleles revealed that mutagenic translesion synthesis, nucleotide excision repair, and homologous recombination are all regulated through residues at the subunit interface. Moreover, genetic characterization of one of our mutants identifies a new sub-branch of nucleotide excision repair. Based on these results we conclude that residues at the subunit boundary of PCNA are not only important for the formation of the trimer structure of PCNA, but they constitute a regulatory protein domain that mediates different DNA damage response pathways, as well.",
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