Functional complementation of xeroderma pigmentosum complementation group E by replication protein A in an in vitro system

被引:61
作者
Kazantsev, A [1 ]
Mu, D [1 ]
Nichols, AF [1 ]
Zhao, XD [1 ]
Linn, S [1 ]
Sancar, A [1 ]
机构
[1] UNIV CALIF BERKELEY, DIV BIOCHEM & MOLEC BIOL, BERKELEY, CA 94720 USA
关键词
DNA repair; damage recognition; excision nuclease;
D O I
10.1073/pnas.93.10.5014
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Xeroderma pigmentosum (XP) is caused by a defect in nucleotide excision repair. Patients in the complementation group E (XP-E) have the mildest form of the disease and the highest level of residual repair activity. About 20% of the cell strains derived from XP-E patients lack a damaged DNA-binding protein (DDB) activity that binds to ultraviolet-induced (6-4) photoproducts with high affinity. We report here that cell-free extracts prepared from XP-E cell strains that either lacked or contained DDB activity were severely defective in excising DNA damage including (6-4) photoproducts. However, this excision activity defect was not restored by addition of purified DDB that. In fact, inhibited removal of (6-4) photoproducts by the human excision nuclease reconstituted from purified proteins. Extensive purification of correcting activity from HeLa cells revealed that the correcting activity is inseparable from the human replication/repair protein A [RPA (also known as human single stranded DNA binding protein, HSSB)]. Indeed, supplementing XP-E extracts with recombinant human RPA purified from Escherichia coli restored excision activity. However, no mutation was found in the genes encoding the three subunits of RPA in an XP-E (DDB-) cell line. It is concluded that RPA functionally complements XP-E extracts in vitro, but it is not genetically altered in XP-E patients.
引用
收藏
页码:5014 / 5018
页数:5
相关论文
共 40 条
  • [1] XERODERMA PIGMENTOSUM GROUP-E CELLS LACK A NUCLEAR FACTOR THAT BINDS TO DAMAGED DNA
    CHU, G
    CHANG, E
    [J]. SCIENCE, 1988, 242 (4878) : 564 - 567
  • [2] Cleaver J.E., 1989, The Metabolic Basis of Inherited Disease, VII, P2949
  • [3] REQUIREMENT FOR THE REPLICATION PROTEIN SSB IN HUMAN DNA EXCISION REPAIR
    COVERLEY, D
    KENNY, MK
    MUNN, M
    RUPP, WD
    LANE, DP
    WOOD, RD
    [J]. NATURE, 1991, 349 (6309) : 538 - 541
  • [4] A ROLE FOR THE HUMAN SINGLE-STRANDED-DNA BINDING-PROTEIN HSSB/RPA IN AN EARLY STAGE OF NUCLEOTIDE EXCISION REPAIR
    COVERLEY, D
    KENNY, MK
    LANE, DP
    WOOD, RD
    [J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (15) : 3873 - 3880
  • [5] CHROMOSOMAL LOCALIZATION AND CDNA CLONING OF THE GENES (DDB1 AND DDB2) FOR THE P127 AND P48 SUBUNITS OF A HUMAN DAMAGE-SPECIFIC DNA-BINDING PROTEIN
    DUALAN, R
    BRODY, T
    KEENEY, S
    NICHOLS, AF
    ADMON, A
    LINN, S
    [J]. GENOMICS, 1995, 29 (01) : 62 - 69
  • [6] ERDILE LF, 1991, J BIOL CHEM, V266, P12090
  • [7] ERDILE LF, 1990, J BIOL CHEM, V265, P3177
  • [8] CELLULAR FACTORS REQUIRED FOR MULTIPLE STAGES OF SV40 DNA-REPLICATION INVITRO
    FAIRMAN, MP
    STILLMAN, B
    [J]. EMBO JOURNAL, 1988, 7 (04) : 1211 - 1218
  • [9] FRIEDBERG EC, 1995, DNA REPAIR MUTAGENES, P191
  • [10] RECONSTITUTION OF YEAST NUCLEOTIDE EXCISION-REPAIR WITH PURIFIED RAD PROTEINS, REPLICATION PROTEIN-A, AND TRANSCRIPTION FACTOR TFIIH
    GUZDER, SN
    HABRAKEN, Y
    SUNG, P
    PRAKASH, L
    PRAKASH, S
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (22) : 12973 - 12976