Abstract
Meiotic recombinations in the proximal region of the mouse major histocompatibility complex (MHC) are clustered within certain segments of chromosome, known as hotspots. In this study, we found that one of such hotspots, previously mapped between the Pb and Ob genes, is located very close to the 3′ end of the Lmp2 gene, which encodes a subunit of a proteolytic proteasome. To analyze the molecular basis of the site specificity of hotspots, we examined the structure of the chromatin around this Lmp2 hotspot and another one located in the MHC class II Eb gene, by monitoring DNase I-hypersensitive sites (DHSSs) of the chromatin. DHSSs were detected at the both hotspots in the somatic cells. In the meiotic cells, DHSS was detected within the Eb hotspot, as previously reported, but not in the Lmp2 hotspot. Thus, open structure of chromatin during meiosis, as monitored by hypersensitivity to DNase I, is not a general feature of mouse recombinational hotspots, contrasting the case of the lower eukaryote, S. cerevisiae, in which hotspots are always associated with DHSSs.
Similar content being viewed by others
References
Alt, F.W., Blackwell, T.K., Depinho, R.A., Reth, M.G., Yancopoulos, G.D. (1986). Regulation of genome rearrangement events during lymphocyte differentiation. Immunol. Rev. 89, 5–30.
Bergman, Y., Rice, D., Grosschedl, R., Baltimore, D. (1984). Two regulatory elements for immunoglobulin k light chain gene expression. Proc. Natl. Acad. sci. USA 81, 5520–5524.
Bryda, E.C., DePari, J.A., Sant’;Angelo, D.B., Murphy, D.B., Passmore, H.C. (1992). Multiple sites of crossing over within the Eb recombinational hotspot in the mouse. Mamm. Genome 2, 123–129.
Cao, L., Alani, E., Kleckner, N. (1990). A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae. Cell 61, 1089–1101.
Cho, S., Attaya, M., Brown, M.G., Monaco, J.J. (1991). A cluster of transcribed sequences between the Pb and Ob genes of the murine major histocompatibility complex. Proc. Natl. Acad. sci. USA 88, 5197–5201.
Church, G.M., Gilbert, W. (1984). Genomic sequencing. Proc. Natl. Acad. sci. USA 81, 1991–1995.
Dietrich, W.F., Miller, J.C., Steen, R.G., Merchant, M., Damron, D., Nahf, R., Gross, A., Joyce, D.C., Wessel, M., Dredge, R.D., Marquis, A., Stein, L.D., Goodman, N., Page, D.C., Lander, E.S. (1994). A genetic map of the mouse with 4,006 simple sequence length polymorphisms. Nature Genet. 7, 220–245.
Falkner, F.G., Zachau, H.G. (1984). Correct transcription of an immuno-globulin k gene requires an upstream fragment containing conserved sequence elements. Nature 310, 71–74.
Feinberg, A.P., Vogelstein, B. (1984). A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 137, 1266–1267.
Fischer Lindahl, K. (1991). His and hers recombinational hotspots. Trends Genet. 7, 273–276.
Grabske, R.J., Lake, S., Gledhill, B.L., Meistrich,.L. (1975). Centrifugal elutriation: separation of spermatogenic cells on the basis of sedimentation velocity. J. Cell. Physiol. 86, 177–190.
Grimm, C, Schaer, P., Munz, P., Kohli, J. (1991). The strong ADH1 promoter stimulates mitotic and meiotic recombination at the ADE6 gene of Schizosaccharomyces pombe. Mol. Cell. Biol. 11, 289–298.
Heine, D., Khambata, S., Wydner, K.S., Passmore, H.C. (1994). Analysis of recombinational hotspots associated with the p haplotype of the mouse MHC. Genomics 23, 168–177.
Heinlein, U.A.O., Lange-Sablitzky, R., Schaal, H., Wille, W. (1986). Molecular characterization of the MT-family of dispersed middle-repetitive DNA in rodent genomes. Nucleic Acids Res. 14, 6403–6416.
Henlkoff, S. (1984). Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28, 351–359.
Klein, J. (1975). Biology of the Mouse Histocompatibility-2 Complex. (New York: Springer-Verlag New York Inc).
Kobori, J.A., Strauss, E., Minard, K., Hood, L. (1986). Molecular analysis of the hotspot of recombination in the murine histocompatibility complex. Science 234, 173–179.
Lafuse, W.P., Berg, N., Savariayan, S., David, C.S. (1986). Mapping of a second recombination hotspot within the I-E region of the mouse H-2 gene complex. J. Exp. Med. 163, 1518–1528.
Lafuse, W.P., Lee, S.T., David, C.S. (1990). Molecular analysis of two recombinant mouse strains with crossovers in the Ea recombination hot spot. J. Immunogenet. 17, 169–176.
Lenardo, M.J., Baltimore, D. (1989). NF-kB: A pleiotropic mediator of inducible and tissue-specific gene control. Cell 58, 227–229.
Ling, X., Shenkar, R., Sakai, D., Arnheim, N. (1993). The mouse Eb meiotic recombination hotspot contains a tissue-specific transcriptional enhancer. Immunogenetics 37, 331–336.
Martinez, C.K., Monaco, J.J. (1991). Homology of proteasome subunits to a major histocompatibility complex-linked LMP gene. Nature 353, 664–667.
Meistrich, M.L., Trostle, P.K. (1975). Separation of mouse testis cells by equilibrium density centrifugation in renografin gradients. Exp. Cell Res. 92, 231–244.
Nicolas, A., Treco, Schultes, N.P., Szostak, J.W. (1989). An initiation site for meiotic gene conversion in the yeast Saccharomyces cerevisiae. Nature 338, 35–39.
Ohta, K., Shibata, T., Nicolas, A. (1994). Changes in chromatin structure at recombination initiation sites during yeast meiosis. EMBO J. 13, 5754–5763.
Petes, T.D., Malone, R.E., Symington, L.S. (1991). Recombination in yeast. In The Molecular and Cellular Biology of the Yeast Saccharomyces: Genome Dynamics, Protein Synthesis, and Energetics, J.R. Broach, J.R. Pringle, E.W. Jones, eds. (Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press), pp. 407–521.
Saha, B.K. (1993). Recruitment of multiple alleles within the Eb recombinational hotspot in murine MHC. Mamm. Genome 4, 565–570.
Sambrook, J., Fritsch, E.F., Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press).
Sanger, F., Nicklen, S., Coulson, A.R. (1977). DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. sci. USA 74, 5463–5467.
Sasaki, H., Jones, P.A., Chaillet, J.R., Fergunson-Smith, A.C., Barton, S.C., Reik, W., Surani, M.A. (1992). Parental imprinting: potentially active chromatin of the repressed maternal allele of the mouse insulin-like growth factor II (Igf2) gene. Genes & Dev. 6, 1843–1856.
Schultes, N.P., Szostak, J.W. (1991). A poly(dA • dT) tract is a component of the recombination initiation site at the ARG4 locus in Saccharomyces cerevisiae. Mol. Cell. Biol. 11, 322–328.
Shenkar, R., Shen, M., Arnheim, N. (1991). DNase I-hypersensitive sites and transcription factor-binding motifs within the mouse Eb meiotic recombination hot spot. Mol. Cell. Biol. 11, 1813–1819.
Shiroishi, T., Sagai, T., Moriwaki, K. (1982). A new wild-derived H-2 haplotype enhancing K-IA recombination. Nature 300, 370–372.
Shiroishi, T., Sagai, T. Moriwaki, K. (1987). Sexual preference of meiotic recombination within the H-2 copmlex. Immunogenetics 25, 258–262.
Shiroishi, T., Hanzawa, N., Sagai, T., Ishiura, M., Gojobori, T., Steinmetz, M., Moriwaki, K. (1990). Recombinational hotspot specific to female meiosis in the mouse major histocompatibility complex. Immunogenetics 31, 79–88.
Shiroishi, T., Sagai, T., Hanzawa, N., Gotoh, H., Moriwaki, K. (1991). Genetic control of sex-dependent meiotic recombination in the major histocompatibility complex of the mouse. EMBO J. 10, 681–686.
Shiroishi, T., Sagai, T., Moriwaki, K. (1993). Hotspots of meiotic recombination in the mouse major histocompatibility complex. Genetica 88, 187–196.
Steinmetz, M., Minard, K., Horvath, S., McNicholas, J., Srelinger, J., Wake, C, Long, E., Mach, B., Hood, L. (1982). A molecular map of the immune response region from the major histocompatibility complex of the mouse. Nature 300, 35–42.
Steinmetz, M., Stephan, D., Fischer-Lindahl, K. (1986). Gene organization and recombinational hotspots in the murine major histocompatibility complex. Cell 44, 895–904.
Sun, H., Treco, D., Schultes, N.P., Szostak, J.W. (1989). Double-strand breaks at an initiation site for meiotic gene conversion. Nature 338, 87–90.
Uematsu, Y., Kiefer, H., Schulze, R., Fischer-Lindahl, K., Steinmetz, M. (1986). Molecular characterization of a meiotic recombinational hotspot enhancing homologous equal crossing-over. EMBO J. 5, 2123–2129.
Wang, A.M., Doyle, M.V., Mark, D.F. (1989). Quantitation of mRNA by the polymerase chain reaction. Proc. Natl. Acad. sci. USA 86, 9717–9721.
White, M.A., Detloff, P., Strand, M., Petes, T.D. (1992). A promoter deletion reduces the rate of mitotic, but not meiotic, recombination at the HIS4 locus in yeast. Curr. Genet. 21, 109–116.
Wu, C. (1989). Analysis of hypersensitive sites in chromatin. Methods Enzymol. 170 269–289.
Wu, T.-C, Lichten, M. (1994). Meiosis-induced double-strand break sites determined by yeast chromatin structure. Science 263, 515–518.
Zenvirth, D., Arbel, T., Sherman, A., Goldway, M., Klein, S., Simchen, G. (1992). Multiple sites for double-strand breaks in whole meiotic chromosomes of Saccharomyces cerevisiae. EMBO J. 11, 3441–3447.
Zhou, P., Cao, H., Smart, M., David, C. (1993). Molecular basis of genetic polymorphism in major histocompatibility complex-linked proteasome gene (Lmp2). Proc. Natl. Acad. sci. USA 90, 2681–2684.
Zimmerer, E.J., Passmore, H.C. (1991). Structural and genetic properties of the Eb recombinational hotspot in the mouse. Immunogenetics 33, 132–140.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mizuno, K., Koide, T., Sagai, T. et al. Molecular analysis of a recombinational hotspot adjacent to Lmp2 gene in the mouse MHC: Fine location and chromatin structure. Mammalian Genome 7, 490–496 (1996). https://doi.org/10.1007/s003359900149
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s003359900149