TY - JOUR
T1 - V(D)J recombination
T2 - In vitro coding joint formation
AU - Weis-Garcia, Frances
AU - Besmer, Eva
AU - Sawchuk, Dennis J.
AU - Yu, Wong
AU - Hu, Yun
AU - Cassard, Sylvanie
AU - Nussenzweig, Michel C.
AU - Cortes, Patricia
PY - 1997/11
Y1 - 1997/11
N2 - Antigen receptor genes are assembled through a mechanism known as V(D)J recombination, which involves two different joining reactions: signal and coding joining. Formation of these joints is essential for antigen receptor assembly as well as maintaining chromosomal integrity. Here we report on a cell-free system for coding joint formation using deletion and inversion recombination substrates. In vitro coding joint formation requires RAG1, RAG2, and heat-labile factors present in the nuclear extract of nonlymphoid cells. Both inversion- and deletion-mediated coding joint reactions produce diverse coding joints, with deletions and P nucleotide addition. We also show that deletion-mediated coding joint formation follows the 12/23 rule and requires the catalytic subunit of DNA-dependent protein kinase.
AB - Antigen receptor genes are assembled through a mechanism known as V(D)J recombination, which involves two different joining reactions: signal and coding joining. Formation of these joints is essential for antigen receptor assembly as well as maintaining chromosomal integrity. Here we report on a cell-free system for coding joint formation using deletion and inversion recombination substrates. In vitro coding joint formation requires RAG1, RAG2, and heat-labile factors present in the nuclear extract of nonlymphoid cells. Both inversion- and deletion-mediated coding joint reactions produce diverse coding joints, with deletions and P nucleotide addition. We also show that deletion-mediated coding joint formation follows the 12/23 rule and requires the catalytic subunit of DNA-dependent protein kinase.
UR - http://www.scopus.com/inward/record.url?scp=0030808573&partnerID=8YFLogxK
U2 - 10.1128/MCB.17.11.6379
DO - 10.1128/MCB.17.11.6379
M3 - Article
C2 - 9343399
AN - SCOPUS:0030808573
VL - 17
SP - 6379
EP - 6385
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
SN - 0270-7306
IS - 11
ER -