title: DNA binding activity studies and computational approach of mutant SRY in patients with 46, XY complete pure gonadal dysgenesis.
abstract: Mutations of SRY are the cause of 46,XY complete pure gonadal dysgenesis (PGD) in 10-15% of patients. In this study, DNA was isolated and sequenced from blood leukocytes and from paraffin-embedded gonadal tissue in five patients with 46,XY complete PGD. DNA binding capability was analyzed by three different methods. The structure of the full length SRY and its mutant proteins was carried out using a protein molecular model. DNA analysis revealed two mutations and one synonymous polymorphism: in patient #4 a Y96C mutation, and a E156 polymorphism; in patient #5 a S143G mosaic mutation limited to gonadal tissue. We demonstrated, by all methods used, that both mutant proteins reduced SRY DNA binding activity. The three-dimensional structure of SRY suggested that besides the HMG box, the carboxy-terminal region of SRY interacts with DNA. In conclusion, we identified two SRY mutations and a polymorphism in two patients with 46,XY complete PGD, demonstrating the importance of the carboxy-terminal region of SRY in DNA binding activity.
authors.affiliation: Instituto Mexicano del Seguro Social, Unidad de Investigación Médica en Biología del Desarrollo, Centro Médico Nacional Siglo XXI, Coordinación de Investigación en Salud, Mexico, D.F., Mexico.
authors.forename: Irene
authors.initials: I
authors.lastname: Sánchez-Moreno
authors.forename: Patricia
authors.initials: P
authors.lastname: Canto
authors.forename: Patricia
authors.initials: P
authors.lastname: Munguía
authors.forename: Mario Bermúdez
authors.initials: MB
authors.lastname: de León
authors.forename: Bulmaro
authors.initials: B
authors.lastname: Cisneros
authors.forename: Felipe
authors.initials: F
authors.lastname: Vilchis
authors.forename: Edgardo
authors.initials: E
authors.lastname: Reyes
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Méndez
mesh_terms: D001483:Base Sequence; D019295:Computational Biology; D004247:DNA / Q000378:metabolism*; D004252:DNA Mutational Analysis; D006061:Gonadal Dysgenesis, 46,XY / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D051876:Sex-Determining Region Y Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D050505:Mutant Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; D004247:DNA