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Mol. Hum. Reprod. Advance Access originally published online on May 2, 2008
Molecular Human Reproduction 2008 14(6):325-330; doi:10.1093/molehr/gan021
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© The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Full-length SRY protein is essential for DNA binding

I. Sánchez-Moreno1,{dagger}, R. Coral-Vázquez2, J.P. Méndez1,3 and P. Canto1,4

1Unidad de Investigación Médica en Biología del Desarrollo, Centro Médico Nacional Siglo XXI, Coordinación de Investigación en Salud, Instituto Mexicano del Seguro Social, C.P. 06720 México, D.F., Mexico 2Unidad de Investigación Médica en Genética Humana, Hospital de Pediatría, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, C.P. 06720 México, D.F., Mexico

4 Correspondence address. Tel/Fax: +52-55-5627-69-00; E-mail: ipcanto{at}yahoo.com.mx

SRY directs testicular development. It has been suggested that the only high-mobility group (HMG) box of the SRY is important for the function of this protein; however, other studies have suggested that the N- and C-terminal regions are also involved in this process. Herein, we analysed and compared in vitro the DNA-binding activity of the full-length SRY and three mutants (HMG box alone, N-terminal less and C-terminal less SRY proteins). DNA-binding capability was analysed by mobility shift assays, optical density and dissociation constant by using pure non-fusion SRY proteins. The structure of the full-length SRY was carried out using a protein molecular model. The HMG box SRY alone and C-terminal less SRY proteins had a statistically diminished DNA binding in comparison with the full-length SRY. In contrast, the affinity for DNA of the N-terminal less SRY was relatively similar to the full-length SRY. Likewise, three-dimensional structure of the full-length SRY suggested that some residues of the C-terminal region of the SRY interact with DNA. We demonstrate the importance that full-length SRY has, particularly the C-terminal region of the protein, in DNA binding in vitro. Likewise, the affinity of the HMG box alone is clearly reduced when compared with the full-length SRY.

Key words: DNA binding/full-length SRY/HMG box SRY/N-terminal less SRY/C-terminal less SRY

{dagger} This work was submitted in partial fulfillment of the requirements for the D.Sc. degree of Sánchez-Moreno I, for obtaining her degree in the Doctorado en Ciencias Biomédicas. Universidad Nacional Autónoma de México.

3 Present address: Unidad de Investigación en Obesidad: Facultad de Medicina, Universidad Nacional Autónoma de México; Clínica de Obesidad, Instituto Nacional de Ciencias Médicas y Nutrición ‘Salvador Zubirán’, México, D.F., Mexico

Submitted on February 14, 2008; resubmitted on April 14, 2008; accepted on April 16, 2008.


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