Abstract
The X-linked immunodeficiency (Xid) in CBA/N mice serves as a model for the X-linked agammaglobulinemia (XLA) syndrome in man. X-chromosome inactivation in F1 heterozygotes derived from CBA/N (Xxid/Xxid) and B6.Pgk-1a (X +/Y) was investigated by monitoring the methylation status of the individualPgk-1 alleles,Pgk-1b andPgk-1a, respectively, using a novel Tth111I RFLP. Results indicate that in circulating B lymphocytes of female heterozygotes, only the X chromosomes carrying the normal alleles(X +) are active (nonrandom inactivation of the X chromosome), whereas in non-B cells both the X chromosomes (X+ and Xxid) are active (random inactivation of the X chromosome). These results were further confirmed by direct evaluation of transcription of theBtk gene, the gene mutated both in Xid and in XLA.
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References
Adra, C. N., Boer, P. H., and McBurney, M. W. (1987). Cloning and expression of the mousePgk-1 gene and the nucleotide sequence of its promoter.Gene 6065.
Avner, P., Amar, L., Dandolo, L., and Guenet, J. L. (1988). Genetic analysis of the mouse using interspecific crosses.Trends Genet. 418.
Boer, P. H., Potten, H., Adra, C. N., Jardine, K., Mullhofer, G., and McBurney, M. W. (1990). Polymorphisms in the coding and noncoding regions of murinePgk-1 alleles.Biochem. Genet. 28299.
Bruton, O. C. (1952). Agammaglobulinemia.Pediatrics 9722.
Cedar, H. (1988). DNA methylation and gene activity.Cell 533.
Chomczynski, P., and Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.Anal. Biochem. 162156.
Cohen, P. L., Scher, I., and Mosier, D. E. (1976). In vitro studies of the genetically determined unresponsiveness to thymus-independent antigens in CBA/N mice.J. Immunol. 116301.
Dailey, M. O., and Hunter, R. L. (1977). Induction of cell-mediated immunity to chemically modified antigens in guinea pigs. I. Characterization of the immune response to lipid-conjugated protein antigens.J. Immunol. 118957.
de Weers, M., Verschuren, M. C. M., Kraakman, M. E. M., Mensink, R. G. J., Schuurman, R. K. B., van Dongen, J. J. M., and Hendriks, R. W. (1993). The Bruton's tyrosine kinase gene is expressed throughout B cell differentiation, from early precursor B cell stages preceding immunoglobulin gene rearrangement up to mature B cell stages.Eur. J. Immunol. 233109.
Fearon, E. R., Winkelstein, J. A., Civin, C. L., Pardoll, D. M., and Vogelstein, B. (1987). Carrier detection in X-linked agammaglobulinemia by analysis of X-chromosome inactivation.N. Engl. J. Med. 316427.
Fernandez, C., and Moller, G. (1977). Immunological unresponsiveness to thymus-independent antigens: Two fundamentally different genetic mechanisms of B cell unresponsiveness to dextran.J. Exp. Med. 1461663.
Finkelman, F. D., Smith, A. H., Scher, I., and Paul, W. E. (1975). Abnormal ratio of membrane immunoglobulin classes in mice with an X-linked B lymphocyte defect.J. Exp. Med. 1421316.
Forrester, L. M., Ansell, J. D., and Micklem, H. S. (1987). Development of B lymphocytes in mice heterozygous for the X-linked immunodeficiency (xid) mutation.J. Exp. Med. 165949.
Fowlis, D. J., Ansell, J. D., and Micklem, H. S. (1991). Further evidence for the importance of parental source of the Xce allele in X chromosome inactivation.Genet. Res. 5863.
Genevier, H. C., Hinshelwood, S., Gaspar, H. B., Rigley, K. P., Brown, D., Saeland, S., Rousset, F., Levinsky, R. J., Callard, R. E., Kinnon, C., and Lovering, R. C. (1994). Expression of Bruton's tyrosine kinase protein within the B cell lineage.Eur. J. Immunol. 243100.
Gilliland, D. G., Blanchard, K. L., Levy, J., Perrin, S., and Bunn, F. (1991). Clonality in myeloproliferative disorders: Analysis by means of polymerase chain reaction.Proc. Natl. Acad. Sci (USA) 886848.
Green, M. C. (1990). Catalog of mutant genes and polymorphic loci. In Lyon, M. F., and Searle, A. G. (eds.),Genetic Variants and Strains of the Laboratory Mouse Oxford University Press, Oxford, UK, p. 391.
Mage, M. G., McHugh, L. L., and Rothstein, T. L. (1977). Mouse lymphocytes with and without surface immunoglobulin: Preparative scale separation in polytyrene tissue culture dishes coated with specifically purified anti-immunoglobulin.J. Immunol. Methods 1547.
Monk, M. (1986). Methylation and the X chromosome.BioEssays 4204.
Nahm, M. H., Paslay, J. W., and Davie, J. M. (1983). Unbalanced X chromosome mosaicism in B cells of mice with X-linked immunodeficiency.J. Exp. Med. 158920.
Nielsen, J. T., and Chapman, V. M. (1977). Electrophoretic variation for X-chromosome linked phosphoglycerate kinase (PGK-1) in mouse.Genetics 87319.
Perlmutter, R. M., Nahm, M., Stein, K. E., Slack, J., Zitron, I., Paul, W. E., and Davie, J. M. (1979). Immunoglobulin subclass-specific immunodeficiency in mice with an X-linked B-lymphocyte defect.J. Exp. Med. 149993.
Rawlings, D. J., Saffran, D. C., Tsukada, S., Largaespada, D. A., Grimaldi, J. C., Cohen, L., Mohr, R. N., Bazan, J. F., Howard, M., Copeland, N. G., Jenkins, N. A., and Witte, O. N. (1993). Mutation of unique region of Bruton's tyrosine kinase in immunodeficient XID mice.Science 261358.
Rawlings, D. J., and Witte, O. N. (1994). Bruton's tyrosine kinase is a key regulator in B-cell development.Immunol. Rev. 138105.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Scher, I. (1982). The CBA/N mouse strain: An experimental model illustrating the influence of the X chromosome on immunity.Adv. Immunol. 331.
Scher, I., Berning, A. K., Kessler, S., and Finkelman, F. D. (1980). Development of B lymphocytes in the mouse: Studies of the frequency and distribution of surface IgM and IgD in normal and immune-defective CBA/N F1 mice.J. Immunol. 1251686.
Singer-Sam, J., Grant, M., Lebon, J. M., Okayama, K., Chapman, V. M., Monk, M., and Riggs, A. D. (1990). Use of a HpaII-polymerase chain reaction assay to study DNA methylation in thePgk-1 CpG island of mouse embroys at the time of X-chromosome inactivation.Mol. Cell. Biol. 104987.
Smith, C. I. E., Islam, K. B., Vorechovsky, I., Olerup, O., Wallin, E., Rabbani, H., Baskin, B., and Hammarstrom, L. (1994a). X-linked agammaglobulinemia and other immunoglobulin deficiencies.Immunol. Rev. 138159.
Smith, C. I. E., Baskin, B., Humire-Greiff, P., Zhou, J., Olsson, P. G., Maniar, H. S., Kjellen, P., Lambris, J. D., Christensson, B., Hammarstrom, L., Bentley, D., Vetrie, D., Islam, K. B., Vorechovsky, I., and Sideras, P. (1994b). Expression of Bruton's agammaglobulinemia tyrosine kinase gene, Btk, is selectively down-regulated in T lymphocytes and plasma cells.J. Immunol. 152557.
Takayama, K., Olsen, M., Datta, P., and Hunter, R. L. (1991). Adjuvant activity of non-ionic block copolymers. V. Modulation of antibody isotype by lipopolysaccharides, lipid A and precursors.Vaccine 9257.
Thomas, J. D., Sideras, P., Smith, C. I. E., Vorechovsky, I., Chapman, V., and Paul, W. E. (1993). Colocalization of X-linked agammaglobulinemia and X-linked immunodeficiency gene.Science 261355.
Tsukada, S., Saffran, D. C., Rawlings, D. J., Parolini, O., Allen, R. C., Klisak, I., Sparkes, R. S., Kubagawa, H., Mohandas, T., Quan, S., Belmont, J. W., Cooper, M. D., Conley, M. E., and Witte, O. N. (1993). Deficient expression of a B cell cytoplasmic tyrosine kinase in human X-linked agammaglobulinemia.Cell 72279.
van Kamp, H., Jansen, R., Willemze, R., Fibbe, W. F., and Landegent, J. E. (1991). Studies on clonality by PCR analysis of the PGK-1 gene.Nucleic Acids Res. 192794.
Vetrie, D., Vorechovsky, I., Sideras, P., Holland, J., Davies, A., Flinter, F., Hammarstrom, L., Kinnon, C., Levinsky, R., Bobrow, M., Smith, C. I. E., and Bentley, D. R. (1993). The gene involved in X-linked agammaglobulinemia is a member of the SRC family of protein-tyrosine kinases.Nature (London) 361226.
Wicker, L. S., and Scher, I. (1986). X-linked immune deficiency (xid) of CBA/N mice.Curr. Top. Micro. Immunol. 12487.
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Shanmugam, V., Chapman, V.M., Sell, K.W. et al. A novelTth111I restriction fragment length polymorphism (RFLP) allows tracing of X-chromosome inactivation in the (Xid) heterozygote. Biochem Genet 34, 17–29 (1996). https://doi.org/10.1007/PL00020593
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DOI: https://doi.org/10.1007/PL00020593