Evidence for somatic rearrangement of immunoglobulin genes coding for variable and constant regions.

Abstract

A high-molecular-weight DNA from Balb/c mouse early embryo or from MOPC 321 plasmacytoma (a .kappa. chain producer) was digested to completion with Bacillus amyloliquefaciens strain H restriction enzyme (BamH I). The resulting DNA fragments were fractionated according to size in preparative agarose gel electrophoresis. DNA fragments carrying gene sequences coding for the variable or constant region of .kappa. chains were detected by hybridization with purified, 125I-labeled, whole MOPC 321 .kappa. mRNA and with its 3''-end half. The pattern of hybridization was completely different in the genomes of embryo cells and of the plasmacytoma. The pattern of embryo DNA showed 2 components, one of which (MW = 6.0 million) hybridized with C-gene sequences and the other (MW = 3.9 million) with V-gene sequences. The pattern of the tumor DNA showed a single component that hybridized with both V-gene and C-gene sequences and that is smaller (MW = 2.4 million) than either of the components in embryo DNA. V.kappa. and C.kappa. genes, which are some distance away from each other in the embryo cells, are joined to form a contiguous polynucleotide stretch during differentiation of lymphocytes. Such joining occurs in both of the homologous chromosomes. Relevance of these findings with respect to models for V-C gene joining, activation of a specific V.kappa. gene, and allelic exclusion in immunoglobulin gene loci is discussed.