Crystal Structure of SUMO-Modified Proliferating Cell Nuclear Antigen

Bret D Freudenthal; John E Brogie; Lokesh Gakhar; Christine M Kondratick; M. Todd Washington

doi:10.1016/j.jmb.2010.12.015

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Crystal Structure of SUMO-Modified Proliferating Cell Nuclear Antigen

Journal article

Peer reviewed

Crystal Structure of SUMO-Modified Proliferating Cell Nuclear Antigen

Bret D Freudenthal, John E Brogie, Lokesh Gakhar, Christine M Kondratick and M. Todd Washington

Journal of molecular biology, Vol.406(1), pp.9-17

2011

DOI: 10.1016/j.jmb.2010.12.015

PMCID: PMC3193175

PMID: 21167178

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Abstract

Eukaryotic proliferating cell nuclear antigen (PCNA) is a replication accessory protein that functions in DNA replication, repair, and recombination. The various functions of PCNA are regulated by posttranslational modifications including mono-ubiquitylation, which promotes translesion synthesis, and sumoylation, which inhibits recombination. To understand how SUMO modification regulates PCNA, we generated a split SUMO-modified PCNA protein and showed that it supports cell viability and stimulates DNA polymerase δ activity. We then determined its X-ray crystal structure and found that SUMO occupies a position on the back face of the PCNA ring, which is distinct from the position occupied by ubiquitin in the structure of ubiquitin-modified PCNA. We propose that the back of PCNA has evolved to be a site of regulation that can be easily modified without disrupting ongoing reactions on the front of PCNA, such as normal DNA replication. Moreover, these modifications likely allow PCNA to function as a tool belt, whereby proteins can be recruited to the replication machinery via the back of PCNA and be held in reserve until needed. [Display omitted] ► PCNA sumoylation inhibits unwanted DNA recombination during DNA replication. ► We generated a split SUMO-modified PCNA protein that functions in vitro and in vivo. ► We determined the X-ray crystal structure of SUMO-modified PCNA to 2.8 Å resolution. ► The SUMO moiety occupies a position on the back face of the PCNA ring. ► The position of the SUMO suggests that SUMO-modified PCNA may function as a tool belt.

DNA replication

DNA recombination

protein–DNA interactions

DNA repair

translesion synthesis

Details

Title: Subtitle: Crystal Structure of SUMO-Modified Proliferating Cell Nuclear Antigen
Creators: Bret D Freudenthal - Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-1109, USA
John E Brogie - Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-1109, USA
Lokesh Gakhar - Protein Crystallography Facility, University of Iowa College of Medicine, Iowa City, IA 52242, USA
Christine M Kondratick - Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-1109, USA
M. Todd Washington - Department of Biochemistry, University of Iowa College of Medicine, Iowa City, IA 52242-1109, USA
Resource Type: Journal article
Publication Details: Journal of molecular biology, Vol.406(1), pp.9-17
DOI: 10.1016/j.jmb.2010.12.015
PMID: 21167178
PMCID: PMC3193175
NLM abbreviation: J Mol Biol
ISSN: 0022-2836
eISSN: 1089-8638
Publisher: Elsevier Ltd
Grant note: DOI: 10.13039/100000057, name: National Institute of General Medical Sciences
Language: English
Date published: 2011
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology; Medicine Administration
Record Identifier: 9984024405802771

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