A hot spot for RAD51C interactions revealed by a peptide that sensitizes cells to cisplatin

被引:14
作者
Connell, PP [1 ]
Siddiqui, N [1 ]
Hoffman, S [1 ]
Kuang, A [1 ]
Khatipov, EA [1 ]
Weichselbaum, RR [1 ]
Bishop, DK [1 ]
机构
[1] Univ Chicago, Dept Radiat & Cellular Oncol, Cummings Life Sci Ctr, Chicago, IL 60637 USA
关键词
D O I
10.1158/0008-5472.CAN-03-3608
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
DNA repair via the homologous recombination pathway requires the recombinase RAD51 and, in vertabrates, five RAD51 paralogs. The paralogs form two complexes in solution, a XRCC3/RAD51C heterodimer and a RAD51B/RAD51C/RAD51D/XRCC2 heterotetramer. Mutation of any one of the five paralog genes prevents subnuclear assembly of recombinase at damaged sites and renders cells 30-100 fold sensitive to DNA cross-linking drugs. Phage display was used to isolate peptides that bind the paralog XRCC3. Sequences of binding peptides showed similarity to residues 14-25 of RAD51C protein. Point mutations in this region of RAD51C altered its interaction with both XRCC3 and RAD51B in a two-hybrid system. A synthetic peptide composed of residues 14-25 of RAD51C fused to a membrane transduction sequence [protein transduction domain 4 (PTD4)], inhibited subnuclear assembly of RAD51 recombinase, and sensitized Chinese hamster ovary cells to cisplatin when added to growth medium. These results suggest that residues 14-25 of RAD51C contribute to a "hot spot" used in both XRCC3-RAD51C and RAD51B-RAD51C interactions. Peptide-based inhibition of homologous recombination may prove useful for improving the efficacy of existing cancer therapies.
引用
收藏
页码:3002 / 3005
页数:4
相关论文
共 21 条
[1]  
BARTEL PL, 1995, METHOD ENZYMOL, V254, P241
[2]   Xrcc3 is required for assembly of Rad51 complexes in vivo [J].
Bishop, DK ;
Ear, U ;
Bhattacharyya, A ;
Calderone, C ;
Beckett, M ;
Weichselbaum, RR ;
Shinohara, A .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (34) :21482-21488
[3]   CROSS-SENSITIVITY OF GAMMA-RAY-SENSITIVE HAMSTER MUTANTS TO CROSS-LINKING AGENTS [J].
CALDECOTT, K ;
JEGGO, P .
MUTATION RESEARCH, 1991, 255 (02) :111-121
[4]   Role of BRCA2 in control of the RAD51 recombination and DNA repair protein [J].
Davies, AA ;
Masson, JY ;
Mcllwraith, MJ ;
Stasiak, AZ ;
Stasiak, A ;
Venkitaraman, AR ;
West, SC .
MOLECULAR CELL, 2001, 7 (02) :273-282
[5]   Isolation and characterization of RAD51C, a new human member of the RAD51 family of related genes [J].
Dosanjh, MK ;
Collins, DW ;
Fan, WF ;
Lennon, GG ;
Albala, JS ;
Shen, ZY ;
Schild, D .
NUCLEIC ACIDS RESEARCH, 1998, 26 (05) :1179-1184
[6]   A CHINESE-HAMSTER OVARY CELL-LINE HYPERSENSITIVE TO IONIZING-RADIATION AND DEFICIENT IN REPAIR REPLICATION [J].
FULLER, LF ;
PAINTER, RB .
MUTATION RESEARCH, 1988, 193 (02) :109-121
[7]   Mammalian Rad51C contributes to DNA cross-link resistance, sister chromatid cohesion and genomic stability [J].
Godthelp, BC ;
Wiegant, WW ;
van Duijn-Goedhart, A ;
Schärer, OD ;
van Buul, PPW ;
Kanaar, R ;
Zdzienicka, MZ .
NUCLEIC ACIDS RESEARCH, 2002, 30 (10) :2172-2182
[8]  
Ho A, 2001, CANCER RES, V61, P474
[9]  
James P, 1996, GENETICS, V144, P1425
[10]   Region and amino acid residues required for Rad51C binding in the human Xrcc3 protein [J].
Kurumizaka, H ;
Enomoto, R ;
Nakada, M ;
Eda, K ;
Yokoyama, S ;
Shibata, T .
NUCLEIC ACIDS RESEARCH, 2003, 31 (14) :4041-4050