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Molecular and Cellular Biology, November 1998, p. 6253-6264, Vol. 18, No. 11
Department of Microbiology and Department of
Biochemistry and Molecular Biophysics, Columbia University College
of Physicians and Surgeons, New York, New York 10032
Received 16 June 1998/Returned for modification 23 July
1998/Accepted 28 July 1998
The importance of V(D)J recombination for generating diversity in
the immune system is well established, but the mechanisms which
regulate V(D)J recombination are still poorly understood. Although
transcription of unrearranged (germ line) immunoglobulin and T-cell
receptor gene segments often precedes V(D)J recombination and has been
implicated in its control, the actual role of germ line transcripts in
V(D)J recombination is not known. We used a sensitive reverse
transcription-PCR assay to study immunoglobulin VH germ
line transcripts in proB lines from RAG-deficient mice. All 10 VH families analyzed were germ line transcribed, and germ line transcription was found in all of the cell lines examined, indicating that active chromatin was present in the VH
region. However, not all VH families were germ line
transcribed in every cell line, and there was a surprising lack of
uniformity in the number and family distribution of germ line
VH transcripts in individual lines. When V(D)J
recombination was activated by restoration of RAG activity,
recombinational activity of endogenous VH genes for which
germ line transcription was observed could be compared with those of
genes for which it was not observed. This analysis revealed multiple
examples of endogenous VH gene segments which were
rearranged in cells where their germ line transcription was not
detectable prior to RAG expression. Thus, our data provide strong
support for the idea that V-(D)J recombination does not require germ
line transcription of the recombining variable gene segment.
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Evidence that Immunoglobulin VH-DJ Recombination Does
Not Require Germ Line Transcription of the Recombining Variable
Gene Segment
*
Corresponding author. Mailing address: Department of
Microbiology, Columbia University College of Physicians and Surgeons, 701 West 168th St., New York, NY 10032. Phone: (212) 305-3504. Fax:
(212) 305-1468. E-mail: KLC1{at}columbia.edu.
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