Differential Requirement for Proliferating Cell Nuclear Antigen in 5′ and 3′ Nick-directed Excision in Human Mismatch Repair

Shuangli Guo, Steven R. Presnell, Fenghua Yuan, Yanbin Zhang, Liya Gu, Guo Min Li

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Proliferating cell nuclear antigen (PCNA) is involved in mammalian mismatch repair at a step prior to or at mismatch excision, but the molecular mechanism of this process is not fully understood. To examine the role of PCNA in mismatch-provoked and nick-directed excision, orientation-specific mismatch removal of hetero-duplexes with a pre-existing nick was monitored in human nuclear extracts supplemented with the PCNA inhibitor protein p21. We show here that, whereas 3′ nick-directed mismatch excision was completely inhibited by low concentrations of p21 or a p21 C-terminal fusion protein, 5′ nick-directed excision was only partially blocked under the same conditions. No further reduction of the 5′ excision was detected when a much higher concentration of p21 C-terminal protein was used. These results suggest the following. (i) There is a differential requirement for PCNA in 3′ and 5′ nick-directed excision; and (ii) 5′ nick-directed excision is conducted by a manner either dependent on or independent of PCNA. Our in vitro reconstitution experiments indeed identified a 5′ nick-directed excision pathway that is dependent on PCNA, hMutSα, and a partially purified fraction from a HeLa nuclear extract.

Original languageEnglish
Pages (from-to)16912-16917
Number of pages6
JournalJournal of Biological Chemistry
Volume279
Issue number17
DOIs
StatePublished - Apr 23 2004
Externally publishedYes

Fingerprint

DNA Mismatch Repair
Proliferating Cell Nuclear Antigen
Repair
Proteins
Protein C
Fusion reactions
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Differential Requirement for Proliferating Cell Nuclear Antigen in 5′ and 3′ Nick-directed Excision in Human Mismatch Repair. / Guo, Shuangli; Presnell, Steven R.; Yuan, Fenghua; Zhang, Yanbin; Gu, Liya; Li, Guo Min.

In: Journal of Biological Chemistry, Vol. 279, No. 17, 23.04.2004, p. 16912-16917.

Research output: Contribution to journalArticle

Guo, Shuangli ; Presnell, Steven R. ; Yuan, Fenghua ; Zhang, Yanbin ; Gu, Liya ; Li, Guo Min. / Differential Requirement for Proliferating Cell Nuclear Antigen in 5′ and 3′ Nick-directed Excision in Human Mismatch Repair. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 17. pp. 16912-16917.
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