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Immunoglobulin Locus
Address correspondence to Andrew Chess, Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, MA 02142. Phone: 617-258-7748; Fax: 617-258-6505; E-mail: chess{at}wi.mit.edu
Rearrangement of antigen receptor genes generates a vast array of antigen receptors on lymphocytes. The establishment of allelic exclusion in immunoglobulin genes requires differential treatment of the two sequence identical alleles. In the case of the
immunoglobulin locus, changes in chromatin structure, methylation, and replication timing of the two alleles are all potentially involved in regulating rearrangement. Additionally, germline transcription of the locus which precedes rearrangement has been proposed to reflect an opening of the chromatin structure rendering it available for rearrangement. As the initial restriction of rearrangement to one allele is critical to the establishment of allelic exclusion, a key question is whether or not germline transcription at the locus is monoallelic or biallelic. We have used a sensitive reverse transcription-polymerase chain reaction (RT-PCR) assay and an RNA–fluorescence in situ hybridization (FISH) to show that germline transcription of the locus is biallelic in wild-type immature B cells and in recombination activating gene (RAG)-/-, µ+ B cells. Therefore, germline transcription is unlikely to dictate which allele will be rearranged first and rather reflects a general opening on both alleles that must be accompanied by a mechanism allowing one of the two alleles to be rearranged first.
Key Words: immunoglobulin • germline transcription • allele exclusion
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