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DNA Repair (Amst). 2017 Feb;50:22-35. doi: 10.1016/j.dnarep.2016.12.003. Epub 2016 Dec 31.

PCNA dependent cellular activities tolerate dramatic perturbations in PCNA client interactions.

Author information

1
Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK.
2
Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK.
3
Target Discovery Institute, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK.
4
University of Exeter Medical School, Barrack Road, Exeter, EX2 5DW, UK.
5
Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK; School of Life Sciences, University of Sussex, Falmer, Brighton, BN1 9RH, UK.
6
Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK. Electronic address: catherine.green@well.ox.ac.uk.

Abstract

Proliferating cell nuclear antigen (PCNA) is an essential cofactor for DNA replication and repair, recruiting multiple proteins to their sites of action. We examined the effects of the PCNAS228I mutation that causes PCNA-associated DNA repair disorder (PARD). Cells from individuals affected by PARD are sensitive to the PCNA inhibitors T3 and T2AA, showing that the S228I mutation has consequences for undamaged cells. Analysis of the binding between PCNA and PCNA-interacting proteins (PIPs) shows that the S228I change dramatically impairs the majority of these interactions, including that of Cdt1, DNMT1, PolD3p66 and PolD4p12. In contrast p21 largely retains the ability to bind PCNAS228I. This property is conferred by the p21 PIP box sequence itself, which is both necessary and sufficient for PCNAS228I binding. Ubiquitination of PCNA is unaffected by the S228I change, which indirectly alters the structure of the inter-domain connecting loop. Despite the dramatic in vitro effects of the PARD mutation on PIP-degron binding, there are only minor alterations to the stability of p21 and Cdt1 in cells from affected individuals. Overall our data suggests that reduced affinity of PCNAS228I for specific clients causes subtle cellular defects in undamaged cells which likely contribute to the etiology of PARD.

KEYWORDS:

DNA repair; DNA replication; PCNA; PCNA-associated repair disorder (PARD)

PMID:
28073635
PMCID:
PMC5264654
DOI:
10.1016/j.dnarep.2016.12.003
[Indexed for MEDLINE]
Free PMC Article

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