TY - JOUR
T1 - In vitro V(D)J recombination
T2 - Signal joint formation
AU - Cortes, Patricia
AU - Weis-Garcia, Frances
AU - Misulovin, Ziva
AU - Nussenzweig, Andre
AU - Lai, Jiann Shiun
AU - Li, Gloria
AU - Nussenzweig, Michel C.
AU - Baltimore, David
N1 - Funding Information:
The present research was financially supported by National Key Technology Research and Development Program of the Ministry of Science and Technology of China (No. 2012BAE04B01), High Technology Research and Development Program of China (No. 2015AA034301), National Natural Science Foundation of China(No.51304041), China Postdoctoral Science Foundation(No. 2013M530936) and Program for New Century Excellent Talents in University(No. N130502001).
PY - 1996/11/26
Y1 - 1996/11/26
N2 - The first step V(D)J recombination, specific cleavage at the recombination signal sequence (RSS), can be carried out by the recombination activating proteins RAG1 and RAG2. In vivo, the cleaved coding and signal ends must be rejoined to generate functional antigen receptors and maintain chromosomal integrity. We have investigated signal joint formation using deletion and inversion substrates in a cell free system. RAG1 and RAG2 alone or in combination were unable to generate signal joints. However, RAG1 and RAG2 complemented with nuclear extracts were able to recombine an extrachromosomal substrate and form precise signal joints. The in vitro reaction resembled authentic V(D)J recombination in being Ku-antigen- dependent.
AB - The first step V(D)J recombination, specific cleavage at the recombination signal sequence (RSS), can be carried out by the recombination activating proteins RAG1 and RAG2. In vivo, the cleaved coding and signal ends must be rejoined to generate functional antigen receptors and maintain chromosomal integrity. We have investigated signal joint formation using deletion and inversion substrates in a cell free system. RAG1 and RAG2 alone or in combination were unable to generate signal joints. However, RAG1 and RAG2 complemented with nuclear extracts were able to recombine an extrachromosomal substrate and form precise signal joints. The in vitro reaction resembled authentic V(D)J recombination in being Ku-antigen- dependent.
KW - recombination activating protein RAG1
KW - recombination activating protein RAG2
UR - https://www.scopus.com/pages/publications/0030477688
U2 - 10.1073/pnas.93.24.14008
DO - 10.1073/pnas.93.24.14008
M3 - Article
C2 - 8943051
AN - SCOPUS:0030477688
SN - 0027-8424
VL - 93
SP - 14008
EP - 14013
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 24
ER -