Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1986 Jan;83(1):135–139. doi: 10.1073/pnas.83.1.135

Cloning an expressed gene shared by the human sex chromosomes.

S M Darling, G S Banting, B Pym, J Wolfe, P N Goodfellow
PMCID: PMC322806  PMID: 2934738

Abstract

The existence of genes shared by mammalian sex chromosomes has been predicted on both evolutionary and functional grounds. However, the only experimental evidence for such genes in humans is the cell-surface antigen encoded by loci on the X and Y chromosomes (MIC2X and MIC2Y, respectively), which is recognized by the monoclonal antibody 12E7. Using the bacteriophage lambda gt11 expression system in Escherichia coli and immunoscreening techniques, we have isolated a cDNA clone whose primary product is recognized by 12E7. Southern blot analysis using somatic cell hybrids containing only the human X or Y chromosomes shows that the sequences reacting with the cDNA clone are localized to the sex chromosomes. In addition, the clone hybridizes to DNAs isolated from mouse cells that have been transfected with human DNA and selected for 12E7 expression on the fluorescence-activated cell sorter. We conclude that the cDNA clone encodes the 12E7 antigen, which is the primary product of the MIC2 loci. The clone was used to explore sequence homology between MIC2X and MIC2Y; these loci are closely related, if not identical.

Full text

PDF
135

Images in this article

136Image
on p.136
136Image
on p.136
137Image
on p.137
137Image
on p.137
137Image
on p.137
137Image
on p.137
137Image
on p.137

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banting G. S., Pym B., Goodfellow P. N. Biochemical analysis of an antigen produced by both human sex chromosomes. EMBO J. 1985 Aug;4(8):1967–1972. doi: 10.1002/j.1460-2075.1985.tb03879.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Batteiger B., Newhall W. J., 5th, Jones R. B. The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 1982 Dec 30;55(3):297–307. doi: 10.1016/0022-1759(82)90089-8. [DOI] [PubMed] [Google Scholar]
  3. Bishop C. E., Guellaen G., Geldwerth D., Voss R., Fellous M., Weissenbach J. Single-copy DNA sequences specific for the human Y chromosome. Nature. 1983 Jun 30;303(5920):831–832. doi: 10.1038/303831a0. [DOI] [PubMed] [Google Scholar]
  4. Bodger M. P., Francis G. E., Delia D., Granger S. M., Janossy G. A monoclonal antibody specific for immature human hemopoietic cells and T lineage cells. J Immunol. 1981 Dec;127(6):2269–2274. [PubMed] [Google Scholar]
  5. Burgoyne P. S. Genetic homology and crossing over in the X and Y chromosomes of Mammals. Hum Genet. 1982;61(2):85–90. doi: 10.1007/BF00274192. [DOI] [PubMed] [Google Scholar]
  6. Chandley A. C., Goetz P., Hargreave T. B., Joseph A. M., Speed R. M. On the nature and extent of XY pairing at meiotic prophase in man. Cytogenet Cell Genet. 1984;38(4):241–247. doi: 10.1159/000132070. [DOI] [PubMed] [Google Scholar]
  7. Cooke H. J., Brown W. A., Rappold G. A. Closely related sequences on human X and Y chromosomes outside the pairing region. Nature. 1984 Sep 20;311(5983):259–261. doi: 10.1038/311259a0. [DOI] [PubMed] [Google Scholar]
  8. Curry C. J., Magenis R. E., Brown M., Lanman J. T., Jr, Tsai J., O'Lague P., Goodfellow P., Mohandas T., Bergner E. A., Shapiro L. J. Inherited chondrodysplasia punctata due to a deletion of the terminal short arm of an X chromosome. N Engl J Med. 1984 Oct 18;311(16):1010–1015. doi: 10.1056/NEJM198410183111603. [DOI] [PubMed] [Google Scholar]
  9. Evans E. P., Burtenshaw M. D., Cattanach B. M. Meitoic crossing-over between the X and Y chromosomes of male mice carrying the sex-reversing (Sxr) factor. Nature. 1982 Dec 2;300(5891):443–445. doi: 10.1038/300443a0. [DOI] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  11. Goodfellow P. N., Tippett P. A human quantitative polymorphism related to Xg blood groups. Nature. 1981 Jan 29;289(5796):404–405. doi: 10.1038/289404a0. [DOI] [PubMed] [Google Scholar]
  12. Goodfellow P., Banting G., Levy R., Povey S., McMichael A. A human X-linked antigen defined by a monoclonal antibody. Somatic Cell Genet. 1980 Nov;6(6):777–787. doi: 10.1007/BF01538976. [DOI] [PubMed] [Google Scholar]
  13. Goodfellow P., Banting G., Sheer D., Ropers H. H., Caine A., Ferguson-Smith M. A., Povey S., Voss R. Genetic evidence that a Y-linked gene in man is homologous to a gene on the X chromosome. Nature. 1983 Mar 24;302(5906):346–349. doi: 10.1038/302346a0. [DOI] [PubMed] [Google Scholar]
  14. Goodfellow P. Expression of the 12E7 antigen is controlled independently by genes on the human X and Y chromosomes. Differentiation. 1983;23 (Suppl):S35–S39. doi: 10.1007/978-3-642-69150-8_6. [DOI] [PubMed] [Google Scholar]
  15. Goodfellow P., Pym B., Mohandas T., Shapiro L. J. The cell surface antigen locus, MIC2X, escapes X-inactivation. Am J Hum Genet. 1984 Jul;36(4):777–782. [PMC free article] [PubMed] [Google Scholar]
  16. KIT S., DUBBS D. R., PIEKARSKI L. J., HSU T. C. DELETION OF THYMIDINE KINASE ACTIVITY FROM L CELLS RESISTANT TO BROMODEOXYURIDINE. Exp Cell Res. 1963 Aug;31:297–312. doi: 10.1016/0014-4827(63)90007-7. [DOI] [PubMed] [Google Scholar]
  17. Kavathas P., Sukhatme V. P., Herzenberg L. A., Parnes J. R. Isolation of the gene encoding the human T-lymphocyte differentiation antigen Leu-2 (T8) by gene transfer and cDNA subtraction. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7688–7692. doi: 10.1073/pnas.81.24.7688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Keitges E., Rivest M., Siniscalco M., Gartler S. M. X-linkage of steroid sulphatase in the mouse is evidence for a functional Y-linked allele. Nature. 1985 May 16;315(6016):226–227. doi: 10.1038/315226a0. [DOI] [PubMed] [Google Scholar]
  19. Klebe R. J., Chen T., Ruddle F. H. Controlled production of proliferating somatic cell hybrids. J Cell Biol. 1970 Apr;45(1):74–82. doi: 10.1083/jcb.45.1.74. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koenig M., Camerino G., Heilig R., Mandel J. L. A DNA fragment from the human X chromosome short arm which detects a partially homologous sequence on the Y chromosomes long arm. Nucleic Acids Res. 1984 May 25;12(10):4097–4109. doi: 10.1093/nar/12.10.4097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Levy R., Dilley J., Fox R. I., Warnke R. A human thymus-leukemia antigen defined by hybridoma monoclonal antibodies. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6552–6556. doi: 10.1073/pnas.76.12.6552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Marcus M., Tantravahi R., Dev V. G., Miller D. A., Miller O. J. Human-mouse cell hybrid with human multiple Y chromosomes. Nature. 1976 Jul 1;262(5563):63–65. doi: 10.1038/262063a0. [DOI] [PubMed] [Google Scholar]
  23. Miller O. J., Drayna D., Goodfellow P. Report of the Committee on the Genetic Constitution of the X and Y Chromosomes. Cytogenet Cell Genet. 1984;37(1-4):176–204. doi: 10.1159/000132009. [DOI] [PubMed] [Google Scholar]
  24. Minowada J., Onuma T., Moore G. E. Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst. 1972 Sep;49(3):891–895. [PubMed] [Google Scholar]
  25. Mohandas T., Shapiro L. J., Sparkes R. S., Sparkes M. C. Regional assignment of the steroid sulfatase-X-linked ichthyosis locus: implications for a noninactivated region on the short arm of human X chromosome. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5779–5783. doi: 10.1073/pnas.76.11.5779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moses M. J., Counce S. J., Paulson D. F. Synaptonemal complex complement of man in spreads of spermatocytes, with details of the sex chromosome pair. Science. 1975 Jan 31;187(4174):363–365. [PubMed] [Google Scholar]
  27. Nabholz M., Miggiano V., Bodmer W. Genetic analysis with human--mouse somatic cell hybrids. Nature. 1969 Jul 26;223(5204):358–363. doi: 10.1038/223358a0. [DOI] [PubMed] [Google Scholar]
  28. Okayama H., Berg P. High-efficiency cloning of full-length cDNA. Mol Cell Biol. 1982 Feb;2(2):161–170. doi: 10.1128/mcb.2.2.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Page D. C., Harper M. E., Love J., Botstein D. Occurrence of a transposition from the X-chromosome long arm to the Y-chromosome short arm during human evolution. Nature. 1984 Sep 13;311(5982):119–123. doi: 10.1038/311119a0. [DOI] [PubMed] [Google Scholar]
  30. Page D., de Martinville B., Barker D., Wyman A., White R., Francke U., Botstein D. Single-copy sequence hybridizes to polymorphic and homologous loci on human X and Y chromosomes. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5352–5356. doi: 10.1073/pnas.79.17.5352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Perucho M., Hanahan D., Wigler M. Genetic and physical linkage of exogenous sequences in transformed cells. Cell. 1980 Nov;22(1 Pt 1):309–317. doi: 10.1016/0092-8674(80)90178-6. [DOI] [PubMed] [Google Scholar]
  32. Schneider U., Schwenk H. U., Bornkamm G. Characterization of EBV-genome negative "null" and "T" cell lines derived from children with acute lymphoblastic leukemia and leukemic transformed non-Hodgkin lymphoma. Int J Cancer. 1977 May 15;19(5):621–626. doi: 10.1002/ijc.2910190505. [DOI] [PubMed] [Google Scholar]
  33. Shapiro L. J., Mohandas T., Weiss R., Romeo G. Non-inactivation of an x-chromosome locus in man. Science. 1979 Jun 15;204(4398):1224–1226. doi: 10.1126/science.156396. [DOI] [PubMed] [Google Scholar]
  34. Singh L., Jones K. W. Sex reversal in the mouse (Mus musculus) is caused by a recurrent nonreciprocal crossover involving the x and an aberrant y chromosome. Cell. 1982 Feb;28(2):205–216. doi: 10.1016/0092-8674(82)90338-5. [DOI] [PubMed] [Google Scholar]
  35. Solari A. J. Synaptosomal complexes and associated structures in microspread human spermatocytes. Chromosoma. 1980;81(3):315–337. doi: 10.1007/BF00368145. [DOI] [PubMed] [Google Scholar]
  36. Stanley K. K., Luzio J. P. Construction of a new family of high efficiency bacterial expression vectors: identification of cDNA clones coding for human liver proteins. EMBO J. 1984 Jun;3(6):1429–1434. doi: 10.1002/j.1460-2075.1984.tb01988.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  39. Wickens M. P., Buell G. N., Schimke R. T. Synthesis of double-stranded DNA complementary to lysozyme, ovomucoid, and ovalbumin mRNAs. Optimization for full length second strand synthesis by Escherichia coli DNA polymerase I. J Biol Chem. 1978 Apr 10;253(7):2483–2495. [PubMed] [Google Scholar]
  40. Wigler M., Silverstein S., Lee L. S., Pellicer A., Cheng Y. c., Axel R. Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell. 1977 May;11(1):223–232. doi: 10.1016/0092-8674(77)90333-6. [DOI] [PubMed] [Google Scholar]
  41. Wilkie N. M., Clements J. B., Boll W., Mantei N., Lonsdale D., Weissmann C. Hybrid plasmids containing an active thymidine kinase gene of Herpes simplex virus 1. Nucleic Acids Res. 1979 Oct 25;7(4):859–877. doi: 10.1093/nar/7.4.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Young J. F., Hockmeyer W. T., Gross M., Ballou W. R., Wirtz R. A., Trosper J. H., Beaudoin R. L., Hollingdale M. R., Miller L. H., Diggs C. L. Expression of Plasmodium falciparum circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine. Science. 1985 May 24;228(4702):958–962. doi: 10.1126/science.2988125. [DOI] [PubMed] [Google Scholar]
  43. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES