V(D)J recombination is not activated by demethylation of the kappa locus

被引:30
作者
Cherry, SR
Beard, C
Jaenisch, R
Baltimore, D [1 ]
机构
[1] CALTECH, Pasadena, CA 91125 USA
[2] MIT, Cambridge, MA 02139 USA
[3] Whitehead Inst, Cambridge Ctr 9, Cambridge, MA 02142 USA
关键词
D O I
10.1073/pnas.150218497
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
V(D)J recombination is thought to be regulated by changes in the accessibility of target sites, such as modulation of methylation. To test whether demethylation of the kappa locus can activate recombination, we generated two recombinationally active B cell lines in which the gene for maintenance of genomic DNA methylation, Dnmt1, was flanked with loxP sites. Transduction with a retrovirus expressing both the cre recombinase and green fluorescent protein allowed us to purify recombinationally active cells devoid of methylation. Loss of methylation of the kappa locus was not sufficient to activate recombination. although transcription was activated in one line. It appears that demethylation of the kappa locus is not the rate-limiting step for altering accessibility and thus regulated demethylation does not generate specificity of recombination.
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收藏
页码:8467 / 8472
页数:6
相关论文
共 40 条
[1]   ORGANIZATION AND REORGANIZATION OF IMMUNOGLOBULIN GENES IN A-MULV TRANSFORMED-CELLS - REARRANGEMENT OF HEAVY BUT NOT LIGHT CHAIN GENES [J].
ALT, F ;
ROSENBERG, N ;
LEWIS, S ;
THOMAS, E ;
BALTIMORE, D .
CELL, 1981, 27 (02) :381-390
[2]   DEVELOPMENT OF THE PRIMARY ANTIBODY REPERTOIRE [J].
ALT, FW ;
BLACKWELL, TK ;
YANCOPOULOS, GD .
SCIENCE, 1987, 238 (4830) :1079-1087
[3]   RECOMBINATION BETWEEN IMMUNOGLOBULIN VARIABLE REGION GENE SEGMENTS IS ENHANCED BY TRANSCRIPTION [J].
BLACKWELL, TK ;
MOORE, MW ;
YANCOPOULOS, GD ;
SUH, H ;
LUTZKER, S ;
SELSING, E ;
ALT, FW .
NATURE, 1986, 324 (6097) :585-589
[4]   A RETROVIRAL PROVIRUS CLOSELY ASSOCIATED WITH THE REN-2 GENE OF DBA/2 MICE [J].
BURT, DW ;
REITH, AD ;
BRAMMAR, WJ .
NUCLEIC ACIDS RESEARCH, 1984, 12 (22) :8579-8593
[5]   TCR-BETA AND TCR-ALPHA GENE ENHANCERS CONFER TISSUE-SPECIFICITY AND STAGE-SPECIFICITY ON V(D)J RECOMBINATION EVENTS [J].
CAPONE, M ;
WATRIN, F ;
FERNEX, C ;
HORVAT, B ;
KRIPPL, B ;
WU, L ;
SCOLLAY, R ;
FERRIER, P .
EMBO JOURNAL, 1993, 12 (11) :4335-4346
[6]   MUTATIONS OF THE INTRONIC IGH ENHANCER AND ITS FLANKING SEQUENCES DIFFERENTIALLY AFFECT ACCESSIBILITY OF THE J(H)-LOCUS [J].
CHEN, JZ ;
YOUNG, F ;
BOTTARO, A ;
STEWART, V ;
SMITH, RK ;
ALT, FW .
EMBO JOURNAL, 1993, 12 (12) :4635-4645
[7]   Chromatin remodeling directly activates V(D)J recombination [J].
Cherry, SR ;
Baltimore, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (19) :10788-10793
[8]   A STRAIN-SPECIFIC MODIFIER ON MOUSE CHROMOSOME-4 CONTROLS THE METHYLATION OF INDEPENDENT TRANSGENE LOCI [J].
ENGLER, P ;
HAASCH, D ;
PINKERT, CA ;
DOGLIO, L ;
GLYMOUR, M ;
BRINSTER, R ;
STORB, U .
CELL, 1991, 65 (06) :939-947
[9]   Nucleosome structure completely inhibits in vitro cleavage by the V(D)J recombinase [J].
Golding, A ;
Chandler, S ;
Ballestar, E ;
Wolffe, AP ;
Schlissel, MS .
EMBO JOURNAL, 1999, 18 (13) :3712-3723
[10]   METHYLATION STATUS OF IMMUNOGLOBULIN CHI GENE SEGMENTS CORRELATES WITH THEIR RECOMBINATION POTENTIAL [J].
GOODHARDT, M ;
CAVELIER, P ;
DOYEN, N ;
KALLENBACH, S ;
BABINET, C ;
ROUGEON, F .
EUROPEAN JOURNAL OF IMMUNOLOGY, 1993, 23 (08) :1789-1795