Identification of two functional PCNA-binding domains in human DNA polymerase κ

Jung Hoon Yoon, Narottam Acharya, Jeseong Park, Debashree Basu, Satya Prakash, Louise Prakash

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Previously, we have shown that human DNA polymerase (Pol) η has two functional PCNA-binding motifs, PIP1 and PIP2, and that a C-terminal deletion of Polη that lacks the ubiquitin-binding UBZ domain and the PIP2 domain but retains the PIP1 domain promotes normal levels of translesion synthesis (TLS) opposite a cis-syn TT dimer in human cells. Here, we identify two PIP domains in Polκ and show that TLS occurs normally in human fibroblast cells in which the pip1 or pip2 mutant Polκ is expressed, but mutational inactivation of both PIP domains renders Polκ nonfunctional in TLS opposite the thymine glycol lesion. Thus, the two PIP domains of Polκ function redundantly in TLS opposite this DNA lesion in human cells. However, and surprisingly, whereas mutational inactivation of the PIP1 domain completely inhibits the stimulation of DNA synthesis by Polκ in the presence of proliferating cell nuclear antigen (PCNA), replication factor C, and replication protein A, mutations in PIP2 have no adverse effect on PCNA-dependent DNA synthesis. This raises the possibility that activation of Polκ PIP2 as a PCNA-binding domain occurs during TLS in human cells and that protein-protein interactions and post-transcriptional modifications are involved in such activation.

Original languageEnglish (US)
Pages (from-to)594-601
Number of pages8
JournalGenes to Cells
Volume19
Issue number7
DOIs
StatePublished - 2014

Fingerprint

Proliferating Cell Nuclear Antigen
DNA-Directed DNA Polymerase
Replication Protein C
Replication Protein A
DNA
Ubiquitin
Proteins
Fibroblasts
Mutation

ASJC Scopus subject areas

  • Genetics
  • Cell Biology
  • Medicine(all)

Cite this

Identification of two functional PCNA-binding domains in human DNA polymerase κ. / Yoon, Jung Hoon; Acharya, Narottam; Park, Jeseong; Basu, Debashree; Prakash, Satya; Prakash, Louise.

In: Genes to Cells, Vol. 19, No. 7, 2014, p. 594-601.

Research output: Contribution to journalArticle

Yoon, Jung Hoon ; Acharya, Narottam ; Park, Jeseong ; Basu, Debashree ; Prakash, Satya ; Prakash, Louise. / Identification of two functional PCNA-binding domains in human DNA polymerase κ. In: Genes to Cells. 2014 ; Vol. 19, No. 7. pp. 594-601.
@article{1ca271c11cce42d68b0dd2b3f6d7f92f,
title = "Identification of two functional PCNA-binding domains in human DNA polymerase κ",
abstract = "Previously, we have shown that human DNA polymerase (Pol) η has two functional PCNA-binding motifs, PIP1 and PIP2, and that a C-terminal deletion of Polη that lacks the ubiquitin-binding UBZ domain and the PIP2 domain but retains the PIP1 domain promotes normal levels of translesion synthesis (TLS) opposite a cis-syn TT dimer in human cells. Here, we identify two PIP domains in Polκ and show that TLS occurs normally in human fibroblast cells in which the pip1 or pip2 mutant Polκ is expressed, but mutational inactivation of both PIP domains renders Polκ nonfunctional in TLS opposite the thymine glycol lesion. Thus, the two PIP domains of Polκ function redundantly in TLS opposite this DNA lesion in human cells. However, and surprisingly, whereas mutational inactivation of the PIP1 domain completely inhibits the stimulation of DNA synthesis by Polκ in the presence of proliferating cell nuclear antigen (PCNA), replication factor C, and replication protein A, mutations in PIP2 have no adverse effect on PCNA-dependent DNA synthesis. This raises the possibility that activation of Polκ PIP2 as a PCNA-binding domain occurs during TLS in human cells and that protein-protein interactions and post-transcriptional modifications are involved in such activation.",
author = "Yoon, {Jung Hoon} and Narottam Acharya and Jeseong Park and Debashree Basu and Satya Prakash and Louise Prakash",
year = "2014",
doi = "10.1111/gtc.12156",
language = "English (US)",
volume = "19",
pages = "594--601",
journal = "Genes to Cells",
issn = "1356-9597",
publisher = "Wiley-Blackwell",
number = "7",

}

TY - JOUR

T1 - Identification of two functional PCNA-binding domains in human DNA polymerase κ

AU - Yoon, Jung Hoon

AU - Acharya, Narottam

AU - Park, Jeseong

AU - Basu, Debashree

AU - Prakash, Satya

AU - Prakash, Louise

PY - 2014

Y1 - 2014

N2 - Previously, we have shown that human DNA polymerase (Pol) η has two functional PCNA-binding motifs, PIP1 and PIP2, and that a C-terminal deletion of Polη that lacks the ubiquitin-binding UBZ domain and the PIP2 domain but retains the PIP1 domain promotes normal levels of translesion synthesis (TLS) opposite a cis-syn TT dimer in human cells. Here, we identify two PIP domains in Polκ and show that TLS occurs normally in human fibroblast cells in which the pip1 or pip2 mutant Polκ is expressed, but mutational inactivation of both PIP domains renders Polκ nonfunctional in TLS opposite the thymine glycol lesion. Thus, the two PIP domains of Polκ function redundantly in TLS opposite this DNA lesion in human cells. However, and surprisingly, whereas mutational inactivation of the PIP1 domain completely inhibits the stimulation of DNA synthesis by Polκ in the presence of proliferating cell nuclear antigen (PCNA), replication factor C, and replication protein A, mutations in PIP2 have no adverse effect on PCNA-dependent DNA synthesis. This raises the possibility that activation of Polκ PIP2 as a PCNA-binding domain occurs during TLS in human cells and that protein-protein interactions and post-transcriptional modifications are involved in such activation.

AB - Previously, we have shown that human DNA polymerase (Pol) η has two functional PCNA-binding motifs, PIP1 and PIP2, and that a C-terminal deletion of Polη that lacks the ubiquitin-binding UBZ domain and the PIP2 domain but retains the PIP1 domain promotes normal levels of translesion synthesis (TLS) opposite a cis-syn TT dimer in human cells. Here, we identify two PIP domains in Polκ and show that TLS occurs normally in human fibroblast cells in which the pip1 or pip2 mutant Polκ is expressed, but mutational inactivation of both PIP domains renders Polκ nonfunctional in TLS opposite the thymine glycol lesion. Thus, the two PIP domains of Polκ function redundantly in TLS opposite this DNA lesion in human cells. However, and surprisingly, whereas mutational inactivation of the PIP1 domain completely inhibits the stimulation of DNA synthesis by Polκ in the presence of proliferating cell nuclear antigen (PCNA), replication factor C, and replication protein A, mutations in PIP2 have no adverse effect on PCNA-dependent DNA synthesis. This raises the possibility that activation of Polκ PIP2 as a PCNA-binding domain occurs during TLS in human cells and that protein-protein interactions and post-transcriptional modifications are involved in such activation.

UR - http://www.scopus.com/inward/record.url?scp=84903199968&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84903199968&partnerID=8YFLogxK

U2 - 10.1111/gtc.12156

DO - 10.1111/gtc.12156

M3 - Article

VL - 19

SP - 594

EP - 601

JO - Genes to Cells

JF - Genes to Cells

SN - 1356-9597

IS - 7

ER -