Evidence of a critical architectural function for the RAG proteins in end processing, protection, and joining in V(D)J recombination

被引:58
作者
Tsai, CL
Drejer, AH
Schatz, DG [1 ]
机构
[1] Yale Univ, Sch Med, Howard Hughes Med Inst, Immunobiol Sect, New Haven, CT 06510 USA
[2] Yale Univ, Sch Med, Dept Mol Biophys & Biochem, New Haven, CT 06510 USA
[3] Yale Univ, Sch Med, Dept Genet, New Haven, CT 06510 USA
关键词
RAG1; RAG2; V(D)J recombination; transposition; DNA double strand break;
D O I
10.1101/gad.984502
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
In addition to creating the DNA double strand breaks that initiate V(D)J recombination, the RAG proteins are thought to play a critical role in the joining phase of the reaction. One such role, suggested by in vitro studies, might be to ensure the structural integrity of postcleavage complexes, but the significance of such a function in vivo is unknown. We have identified RAG1 mutants that are proficient in DNA cleavage but defective in their ability to interact with coding ends after cleavage and in the capture of target DNA for transposition. As a result, these mutants exhibit severe defects in hybrid joint formation, hairpin coding end opening, and transposition in vitro, and in V(D)J recombination in vivo. Our results suggest that the RAG proteins have an architectural function in facilitating proper and efficient V(D)J joining, and a protective function in preventing degradation of broken ends prior to joining.
引用
收藏
页码:1934 / 1949
页数:16
相关论文
共 53 条
[1]   Transposition mediated by RAG1 and RAG2 and its implications for the evolution of the immune system [J].
Agrawal, A ;
Eastman, QM ;
Schatz, DG .
NATURE, 1998, 394 (6695) :744-751
[2]   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
[3]   Definition of minimal domains of interaction within the recombination-activating genes 1 and 2 recombinase complex [J].
Aidinis, V ;
Dias, DC ;
Gomez, CA ;
Bhattacharyya, D ;
Spanopoulou, E ;
Santagata, S .
JOURNAL OF IMMUNOLOGY, 2000, 164 (11) :5826-5832
[4]   Identification of two topologically independent domains in RAG1 and their role in macromolecular interactions relevant to V(D)J recombination [J].
Arbuckle, JL ;
Fauss, LJ ;
Simpson, R ;
Ptaszek, LM ;
Rodgers, KK .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (40) :37093-37101
[5]  
Bailin T, 1999, MOL CELL BIOL, V19, P4664
[6]   Hairpin coding end opening is mediated by RAG1 and RAG2 proteins [J].
Besmer, E ;
Mansilla-Soto, J ;
Cassard, S ;
Sawchuk, DJ ;
Brown, G ;
Sadofsky, M ;
Lewis, SM ;
Nussenzweig, MC ;
Cortes, P .
MOLECULAR CELL, 1998, 2 (06) :817-828
[7]   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
[8]   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
[9]   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
[10]  
Eastman QM, 1999, MOL CELL BIOL, V19, P3788