Summary
XY females (n=17) were analysed for mutations in SRY (sex-determining region Y gene), a gene that has recently been equated with the testis determining factor (TDF). SRY sequences were amplified by the polymerase chain reaction (PCR) and analysed by both the single strand conformational polymorphism assay (SSCP) and DNA sequencing. The DNA from two individuals gave altered SSCP patterns; only these two individuals showed any DNA sequence variation. In both cases, a single base change was found, one altering a tryptophan codon to a stop codon, the other causing a glycine to arginine amino acid substitution. These substitutions lie in the high mobility group (HMG)-related box of the SRY protein, a potential DNA-binding domain. The corresponding regions of DNA from the father of one individual and the paternal uncle of the other, were sequenced and found to be normal. Thus, in both cases, sex reversal is associated with de novo mutations in SRY. Combining this data with two previously published reports, a total of 40 XY females have now been analysed for mutations in SRY. The number of de novo mutations in SRY is now doubled to four, adding further strength to the argument that SRY is TDF.
References
Berta P, Hawkins JR, Sinclair AH, Taylor A, Griffiths B, Good-fellow PN, Fellous M (1990) Genetic evidence equating SRY and the testis determining factor. Nature 348:448–450
Casanova J-L, Pannetier C, Jaulin C, Kourilsky P (1990) Optimal conditions for directly sequenced double-stranded PCR products with Sequenase. Nucleic Acids Res 18:4028
Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995
Goodfellow PJ, Nevanlinna HA, Gorman P, Sheer D, Lam G, Goodfellow PN (1989) Assignment of the gene encoding the beta-subunit of the human fibronectin receptor (β-FNR) to chromosome 10p11.2. Ann Hum Genet 53:15–22
Gubbay J, Collignon J, Koopman P, Capel B, Economou A, Munsterberg A, Vivian N, Goodfellow P, Lovell-Badge R (1990) A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature 346:245–250
Jäger RJ, Anvret M, Hall K, Scherer G (1990) A human XY female with a frameshift mutation in SRY, a candidate testis determining gene. Nature 348:452–454
Jantzen H-M, Admon A, Bell SP, Tjian R (1990) Nucleolar transcription factor hUBF contains a DNA-binding motif with homology to HMG proteins. Nature 344:830–836
Kelly M, Burke J, Smith M, Klar A, Beach D (1988) Four matingtype genes control sexual differentiation in the fission yeast. EMBO J 7:1537–1547
Kolodrubetz D, Burgum A (1990) Duplicated NHP6 genes of Saccharomyces cerevisiae encode proteins homologous to bovine high mobility group protein 1. J Biol Chem 265:3234–3249
Koopman P, Munserberg A, Capel B, Vivian N, Lovell-Badge R (1990) Expression of a candidate sex-determining gene during mouse testis differentiation. Nature 348:450–452
Lopez-Galindez C, Lopez JA, Melero JA, Fuente L de la, Marinez C, Ortin J, Perucho M (1988) Analysis of genetic variability and mapping of point mutations in influenza virus by the RNase A mismatch cleavage method. Proc Natl Acad Sci USA 83:6045–6047
Myers RM, Larin Z, Maniatis T (1985) Detection of single base substitutions by ribonuclease cleavage at mismatches in RNA: DNA duplexes. Science 230:1242–1246
Orita M, Suzuki Y, Sekiya T, Hayashi K (1989) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 8:874–879
Orita M, Sekiya T, Hayashi K (1990) DNA sequence polymorphism in Alu repeats. Genomics 8:271–278
Palmer MS, Sinclair A, Berta P, Ellis NA, Goodfellow PN, Fellous M (1989) Genetic evidence that ZFY is not the testis determining factor. Nature 342:937–939
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sinclair AH, Berta P, Palmer MS, Hawkins JR, Griffiths B, Smith M, Foster J, Frischauf A-M, Lovell-Badge R, Goodfellow PN (1990) A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif. Nature 346:240–244
Staben C, Yanofsky C (1990) Neurospora crassa a mating-type region. Proc Natl Acad Sci USA 87:4917–4921
Walker JM, Gooderham K, Hastings JRB, Mayes E, Johns EW (1980) The primary structures of non-histone chromosomal proteins HMG1 and 2. FEBS Lett 122:264–270
Wetering M van de, Oosterwegel M, Dooijes D, Clevers H (1991) Identification and cloning of TCF-1, a T lymphocyte-specific transcription factor containing a sequence-specific HMG box. EMBO J 10:123–132
Wong Z, Wilson V, Jeffreys AJ, Thein S-L (1986) Cloning a selected fragment from a human DNA ‘fingerprint’: isolation of an extremely polymorphic minisatellite. Nucleic Acids Res 14:4605–4616
Wong Z, Wilson V, Patel I, Povey S, Jeffreys AJ (1987) Characterization of a panel of highly variable minisatellites cloned from human DNA. Ann Hum Genet 51:269–288
Wright JM, Dixon GH (1988) Induction by torsional stress of an altered DNA conformation 5′ upstream of the gene for a high mobility group protein from trout and specific binding to flanking sequences by the gene product HMG-T. Biochemistry 27:576–581
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Hawkins, J.R., Taylor, A., Berta, P. et al. Mutational analysis of SRY: nonsense and missense mutations in XY sex reversal. Hum Genet 88, 471–474 (1992). https://doi.org/10.1007/BF00215684
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DOI: https://doi.org/10.1007/BF00215684