Purification and characterization of the immunoglobulin switch sequence-specific endonuclease (Endo-SR) from bovine spleen.
Mature B lymphocytes are able to specifically alter their Ig isotype expression in response to extracellular stimuli via a highly regulated, deletional recombination process called isotype switch recombination. Switch recombination breakpoints predominantly map to large (1-10 kb), G-rich and highly repetitive switch regions that are located directly upstream of immunoglobulin heavy-chain constant region genes. Switch region repeat structures vary considerably both within and between species, but all switch regions contain disproportionate numbers of two pentamer motifs, TGGGN and TGAGC, that are found at or directly adjacent to most analysed switch junctions. We have recently identified an endonuclease activity, Endo SR, that preferentially cleaves TGGGN and TGAGC switch motifs. We have purified the bovine endonuclease activity to homogeneity and have identified a protein with a molecular weight of approximately 32,000 that directly correlates with enzyme activity. As discussed in this report, we have found that murine and bovine Endo-SR are preferentially enriched in lymphoid tissue nuclear extracts and that both enzymes demonstrate highly similar physical and biochemical characteristics. However, each enzyme demonstrates related but distinctive specificities for consensus and degenerate TGGGN and TGAGC switch pentamer motifs.
Lyon
C J
CJ
Department of Molecular, Cell and Developmental Biology and Molecular Biology Institute, University of California at Los Angeles, 90095-1606, U.S.A.
Aguilera
R J
RJ
D000818:Animals; D001483:Base Sequence; D002413:Cations, Divalent; D002417:Cattle; D016384:Consensus Sequence; D004720:Endonucleases / Q000235:genetics*; D005812:Genes, Switch; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D013154:Spleen / Q000201:enzymology; D013379:Substrate Specificity
D002413:Cations, Divalent; D004720:Endonucleases