RAG1 and RAG2 cooperate in specific binding to the recombination signal sequence in vitro

被引:49
作者
Mo, XM [1 ]
Bailin, T [1 ]
Sadofsky, MJ [1 ]
机构
[1] Med Coll Georgia, Inst Mol Med & Genet, Augusta, GA 30912 USA
关键词
D O I
10.1074/jbc.274.11.7025
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
An essential step in the development of the vertebrate immune system is the DNA level rearrangement of the antigen receptor genes. This process, termed "V(D)J recombination," beans with DNA cleavage at the appropriate sites mediated by the two proteins RAG1 and RAG2. We report here that the two proteins cooperate to bind DNA with significantly higher specificity than either protein alone. Gel purification of the triple complex is performed in the absence of any cross-linking agents. Both proteins remain present in the complex, and UV cross-linking using iodouridine-containing probes shows that RAG1 makes close contacts in both the heptamer and nonamer motifs, The two proteins are also shown to associate with each other in the absence of any DNA. These findings refine our understanding of the protein-DNA interactions that accompany cleavage at the recombination signals.
引用
收藏
页码:7025 / 7031
页数:7
相关论文
共 26 条
[1]   RAG1 and RAG2 form a stable postcleavage synaptic complex with DNA containing signal ends in V(D)J recombination [J].
Agrawal, A ;
Schatz, DG .
CELL, 1997, 89 (01) :43-53
[2]   Distinct roles of RAG1 and RAG2 in binding the V(D)J recombination signal sequences [J].
Akamatsu, Y ;
Oettinger, MA .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (08) :4670-4678
[3]   ANALYSIS OF REGIONS OF RAG-2 IMPORTANT FOR V(D)J RECOMBINATION [J].
CUOMO, CA ;
OETTINGER, MA .
NUCLEIC ACIDS RESEARCH, 1994, 22 (10) :1810-1814
[4]   RAG1 mediates signal sequence recognition and recruitment of RAG2 in V(D)J recombination [J].
Difilippantonio, MJ ;
McMahan, CJ ;
Eastman, QM ;
Spanopoulou, E ;
Schatz, DG .
CELL, 1996, 87 (02) :253-262
[5]   Initiation of V(D)J recombination in vitro obeying the 12/23 rule [J].
Eastman, QM ;
Leu, TMJ ;
Schatz, DG .
NATURE, 1996, 380 (6569) :85-88
[6]  
Gellert M, 1997, ADV IMMUNOL, V64, P39, DOI 10.1016/S0065-2776(08)60886-X
[7]   A complex of RAG-1 and RAG-2 proteins persists on DNA after single-strand cleavage at V(D)J recombination signal sequences [J].
Grawunder, U ;
Lieber, MR .
NUCLEIC ACIDS RESEARCH, 1997, 25 (07) :1375-1382
[8]   V(D)J RECOMBINATION - A FUNCTIONAL DEFINITION OF THE JOINING SIGNALS [J].
HESSE, JE ;
LIEBER, MR ;
MIZUUCHI, K ;
GELLERT, M .
GENES & DEVELOPMENT, 1989, 3 (07) :1053-1061
[9]   A stable RAG1-RAG2-DNA complex that is active in V(D)J Cleavage [J].
Hiom, K ;
Gellert, M .
CELL, 1997, 88 (01) :65-72
[10]   Assembly of a 12/23 paired signal complex: A critical control point in V(D)J recombination [J].
Hiom, K ;
Gellert, M .
MOLECULAR CELL, 1998, 1 (07) :1011-1019