Ubiquitylation of RAG-2 by Skp2-SCF links destruction of the V(D)J recombinase to the cell cycle.
The periodic destruction of RAG-2 at the G1-to-S transition couples V(D)J recombination to the G0 and G1 cell cycle phases and coordinates RAG-mediated DNA cleavage with DNA repair by nonhomologous end joining. To define the mechanism by which this occurs, we reproduced cell cycle-dependent regulation of the V(D)J recombinase in a cell-free system. The ubiquitin-proteasomal pathway carries out destruction of RAG-2 in lysates of S phase cells and during S phase in vivo. Remarkably, the Skp2-SCF ubiquitin ligase, which plays a central role in cell cycle regulation through the destruction of p27, mediates ubiquitylation of RAG-2 in vitro and degradation of RAG-2 in vivo. The regulation of antigen receptor gene assembly by Skp2-SCF provides an unexpected and direct mechanistic link between DNA recombination and the cell cycle.
Jiang
Hao
H
Department of Molecular Biology and Genetics, Institute for Cell Engineering, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Chang
Fu-Chung
FC
Ross
Ashley E
AE
Lee
Jihyun
J
Nakayama
Keiichi
K
Nakayama
Keiko
K
Desiderio
Stephen
S
D002453:Cell Cycle / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D016193:G1 Phase; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins; D011995:Recombination, Genetic; D016196:S Phase; D044786:S-Phase Kinase-Associated Proteins / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*; D045567:VDJ Recombinases / Q000378:metabolism*
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C499311:RAG2 protein, human; D044786:S-Phase Kinase-Associated Proteins; D025801:Ubiquitin; C064655:V(D)J recombination activating protein 2; D045567:VDJ Recombinases