The in vivo pattern of binding of RAG1 and RAG2 to antigen receptor loci.
The critical initial step in V(D)J recombination, binding of RAG1 and RAG2 to recombination signal sequences flanking antigen receptor V, D, and J gene segments, has not previously been characterized in vivo. Here, we demonstrate that RAG protein binding occurs in a highly focal manner to a small region of active chromatin encompassing Ig kappa and Tcr alpha J gene segments and Igh and Tcr beta J and J-proximal D gene segments. Formation of these small RAG-bound regions, which we refer to as recombination centers, occurs in a developmental stage- and lineage-specific manner. Each RAG protein is independently capable of specific binding within recombination centers. While RAG1 binding was detected only at regions containing recombination signal sequences, RAG2 binds at thousands of sites in the genome containing histone 3 trimethylated at lysine 4. We propose that recombination centers coordinate V(D)J recombination by providing discrete sites within which gene segments are captured for recombination.
Ji
Yanhong
Y
Department of Immunobiology, Yale University School of Medicine, 300 Cedar Street, Box 208011, New Haven, CT 06520-8011, USA.
Resch
Wolfgang
W
Corbett
Elizabeth
E
Yamane
Arito
A
Casellas
Rafael
R
Schatz
David G
DG
D000818:Animals; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015322:Gene Rearrangement, B-Lymphocyte; D015329:Gene Rearrangement, T-Lymphocyte; D050438:Genes, Immunoglobulin Heavy Chain; D019673:Genes, T-Cell Receptor alpha; D019674:Genes, T-Cell Receptor beta; D018398:Homeodomain Proteins / Q000378:metabolism*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011995:Recombination, Genetic
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D007145:Immunoglobulin kappa-Chains; C499308:Rag2 protein, mouse; C064658:RAG-1 protein
J Immunol. 2008 Feb 15;180(4):2339-46
18250443
Cell. 2008 Dec 12;135(6):1130-42
19070581
Immunity. 1997 Mar;6(3):225-33
9075923
Immunity. 1999 Mar;10(3):313-22
10204487
Nature. 2004 Mar 4;428(6978):88-93
14999286
Immunol Rev. 2003 Aug;194:77-95
12846809
Nat Immunol. 2005 Dec;6(12):1272-9
16286921
Nucleic Acids Res. 2008 Apr;36(6):2060-72
18281312
Mol Cell. 2005 Jun 10;18(6):699-709
15949444
Mol Cell Biol. 1995 Oct;15(10):5657-70
7565717
DNA Repair (Amst). 2006 Sep 8;5(9-10):1246-58
16798110
Proc Natl Acad Sci U S A. 2007 Oct 23;104(43):17046-51
17939999
Immunol Rev. 2004 Aug;200:70-82
15242397
Cell. 2008 Apr 18;133(2):265-79
18423198
Nature. 2007 Sep 27;449(7161):483-6
17898768
Annu Rev Biochem. 2002;71:101-32
12045092
EMBO J. 2002 Aug 1;21(15):4162-71
12145216
Genes Dev. 1999 Dec 1;13(23):3070-80
10601033
PLoS Biol. 2005 Jun;3(6):e181
15898832
Mol Cell. 2007 Jan 12;25(1):15-30
17218268
Mol Cell. 1998 Dec;2(6):829-39
9885570
Science. 1999 Aug 13;285(5430):1080-4
10446057
Immunity. 2007 Oct;27(4):561-71
17936034
Immunogenetics. 2004 Sep;56(6):399-404
15322776
Adv Immunol. 2006;91:45-109
16938538
Nat Immunol. 2007 Jul;8(7):687-94
17579647
DNA Repair (Amst). 2006 Sep 8;5(9-10):1234-45
16793349
Eur J Immunol. 2008 Sep;38(9):2564-72
18792409
Nat Immunol. 2009 Aug;10(8):817-21
19621044
Nat Immunol. 2009 Jun;10(6):655-64
19448632
Mol Cell. 2000 Jan;5(1):97-107
10678172
Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):903-7
17210914
Cell. 1989 Sep 22;58(6):1035-46
2476238
EMBO J. 1999 Jul 1;18(13):3712-23
10393186
Annu Rev Immunol. 2006;24:541-70
16551259
Nature. 2007 Dec 13;450(7172):1106-10
18033247
Cell. 2004 Apr 16;117(2):171-84
15084256
J Immunol. 1994 Apr 1;152(7):3467-75
7511648
Cell. 1996 Jun 14;85(6):887-97
8681383
Nature. 2006 Jul 27;442(7101):466-70
16799570
Cell. 2007 May 18;129(4):823-37
17512414
Mol Cell. 2010 Jan 29;37(2):282-93
20122409
Genes Dev. 1999 Dec 1;13(23):3059-69
10601032
Immunol Rev. 2004 Aug;200:90-114
15242399
Nat Struct Mol Biol. 2009 May;16(5):499-508
19396172
Cell. 1995 Nov 3;83(3):387-95
8521468
Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):4002-7
17360467
Mol Cell. 2007 Sep 7;27(5):842-50
17803947
Mol Cell Biol. 2002 Jan;22(1):69-77
11739723
Mol Cell. 2009 Jun 12;34(5):535-44
19524534