Identification of regulatory factor X as a novel mismatch repair stimulatory factor

Yanbin Zhang, Fenghua Yuan, Daojing Wang, Liya Gu, Guo Min Li

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

This report describes the identification and purification of a novel mismatch repair stimulatory factor from HeLa cell extracts. This activity copurifies with a proliferating cell nuclear antigen-dependent 5′ → 3′DNAexcision activity during several purification steps but is resolved from the excision activity during gel filtration chromatography using Sephacryl S-300. After purification to near homogeneity, the stimulatory factor is associated with three polypeptides with apparent molecular masses of 68, 36, and 30 kDa. Peptide sequencing analysis by tandem mass spectrometry identified the stimulatory factor as the heterotrimeric regulatory factor X (RFX) complex, which regulates transcription of the class II major histocompatibility complex by facilitating histone acetylation and is defective in the human hereditary immunodeficiency syndrome called bare lymphocyte syndrome. This conclusion was confirmed by the facts that purified recombinant RFX stimulates mismatch repair in an in vitro reconstituted mismatch repair system and that depletion of RFX from nuclear extracts or RFX knockdown in cells reduces mismatch repair activity. As expected, RFX knockdown cells display instability in microsatellite sequences. The possible role of RFX in human MMR repair is discussed.

Original languageEnglish
Pages (from-to)12730-12735
Number of pages6
JournalJournal of Biological Chemistry
Volume283
Issue number19
DOIs
StatePublished - May 9 2008

Fingerprint

Factor X
DNA Mismatch Repair
Repair
Purification
Acetylation
Severe Combined Immunodeficiency
Microsatellite Instability
Peptides
Lymphocytes
Proliferating Cell Nuclear Antigen
Molecular mass
Transcription
Tandem Mass Spectrometry
Chromatography
Cell Extracts
Major Histocompatibility Complex
HeLa Cells
Microsatellite Repeats
Histones
Gel Chromatography

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Identification of regulatory factor X as a novel mismatch repair stimulatory factor. / Zhang, Yanbin; Yuan, Fenghua; Wang, Daojing; Gu, Liya; Li, Guo Min.

In: Journal of Biological Chemistry, Vol. 283, No. 19, 09.05.2008, p. 12730-12735.

Research output: Contribution to journalArticle

Zhang, Yanbin ; Yuan, Fenghua ; Wang, Daojing ; Gu, Liya ; Li, Guo Min. / Identification of regulatory factor X as a novel mismatch repair stimulatory factor. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 19. pp. 12730-12735.
@article{7f6f096f97f547bc95cf70282e5808d3,
title = "Identification of regulatory factor X as a novel mismatch repair stimulatory factor",
abstract = "This report describes the identification and purification of a novel mismatch repair stimulatory factor from HeLa cell extracts. This activity copurifies with a proliferating cell nuclear antigen-dependent 5′ → 3′DNAexcision activity during several purification steps but is resolved from the excision activity during gel filtration chromatography using Sephacryl S-300. After purification to near homogeneity, the stimulatory factor is associated with three polypeptides with apparent molecular masses of 68, 36, and 30 kDa. Peptide sequencing analysis by tandem mass spectrometry identified the stimulatory factor as the heterotrimeric regulatory factor X (RFX) complex, which regulates transcription of the class II major histocompatibility complex by facilitating histone acetylation and is defective in the human hereditary immunodeficiency syndrome called bare lymphocyte syndrome. This conclusion was confirmed by the facts that purified recombinant RFX stimulates mismatch repair in an in vitro reconstituted mismatch repair system and that depletion of RFX from nuclear extracts or RFX knockdown in cells reduces mismatch repair activity. As expected, RFX knockdown cells display instability in microsatellite sequences. The possible role of RFX in human MMR repair is discussed.",
author = "Yanbin Zhang and Fenghua Yuan and Daojing Wang and Liya Gu and Li, {Guo Min}",
year = "2008",
month = "5",
day = "9",
doi = "10.1074/jbc.M800460200",
language = "English",
volume = "283",
pages = "12730--12735",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "19",

}

TY - JOUR

T1 - Identification of regulatory factor X as a novel mismatch repair stimulatory factor

AU - Zhang, Yanbin

AU - Yuan, Fenghua

AU - Wang, Daojing

AU - Gu, Liya

AU - Li, Guo Min

PY - 2008/5/9

Y1 - 2008/5/9

N2 - This report describes the identification and purification of a novel mismatch repair stimulatory factor from HeLa cell extracts. This activity copurifies with a proliferating cell nuclear antigen-dependent 5′ → 3′DNAexcision activity during several purification steps but is resolved from the excision activity during gel filtration chromatography using Sephacryl S-300. After purification to near homogeneity, the stimulatory factor is associated with three polypeptides with apparent molecular masses of 68, 36, and 30 kDa. Peptide sequencing analysis by tandem mass spectrometry identified the stimulatory factor as the heterotrimeric regulatory factor X (RFX) complex, which regulates transcription of the class II major histocompatibility complex by facilitating histone acetylation and is defective in the human hereditary immunodeficiency syndrome called bare lymphocyte syndrome. This conclusion was confirmed by the facts that purified recombinant RFX stimulates mismatch repair in an in vitro reconstituted mismatch repair system and that depletion of RFX from nuclear extracts or RFX knockdown in cells reduces mismatch repair activity. As expected, RFX knockdown cells display instability in microsatellite sequences. The possible role of RFX in human MMR repair is discussed.

AB - This report describes the identification and purification of a novel mismatch repair stimulatory factor from HeLa cell extracts. This activity copurifies with a proliferating cell nuclear antigen-dependent 5′ → 3′DNAexcision activity during several purification steps but is resolved from the excision activity during gel filtration chromatography using Sephacryl S-300. After purification to near homogeneity, the stimulatory factor is associated with three polypeptides with apparent molecular masses of 68, 36, and 30 kDa. Peptide sequencing analysis by tandem mass spectrometry identified the stimulatory factor as the heterotrimeric regulatory factor X (RFX) complex, which regulates transcription of the class II major histocompatibility complex by facilitating histone acetylation and is defective in the human hereditary immunodeficiency syndrome called bare lymphocyte syndrome. This conclusion was confirmed by the facts that purified recombinant RFX stimulates mismatch repair in an in vitro reconstituted mismatch repair system and that depletion of RFX from nuclear extracts or RFX knockdown in cells reduces mismatch repair activity. As expected, RFX knockdown cells display instability in microsatellite sequences. The possible role of RFX in human MMR repair is discussed.

UR - http://www.scopus.com/inward/record.url?scp=45149086930&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=45149086930&partnerID=8YFLogxK

U2 - 10.1074/jbc.M800460200

DO - 10.1074/jbc.M800460200

M3 - Article

C2 - 18319249

AN - SCOPUS:45149086930

VL - 283

SP - 12730

EP - 12735

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 19

ER -