NHERF2/SIP-1 interacts with mouse SRY via a different mechanism than human SRY.
In mammals, male sex determination is controlled by the SRY protein, which drives differentiation of the bipotential embryonic gonads into testes by activating the Sertoli cell differentiation program. The morphological effects of SRY are well documented; however, its molecular mechanism of action remains unknown. Moreover, SRY proteins display high sequence variability among mammalian species, which makes protein motifs difficult to delineate. We previously isolated SIP-1/NHERF2 as a human SRY-interacting protein. SIP-1/NHERF2, a PDZ protein, interacts with the C-terminal extremity of the human SRY protein. Here we showed that the interaction of SIP-1/NHERF2 and SRY via the SIP-1/NHERF2 PDZ1 domain is conserved in mice. However, the interaction occurs via a domain that is internal to the mouse SRY protein and involves a different recognition mechanism than human SRY. Furthermore, we show that mouse and human SRY induce nuclear accumulation of the SIP-1/NHERF2 protein in cultured cells. Finally, a transgenic mouse line expressing green fluorescent protein under the control of the mouse Sry promoter allowed us to show that SRY and SIP-1/NHERF2 are co-expressed in the nucleus of pre-Sertoli cells during testis determination. Taken together, our results suggested that the function of SIP-1/NHERF2 as an SRY cofactor during testis determination is conserved between human and mouse.
Thevenet
Laurie
L
Institut de Génétique Humaine CNRS UPR1142, 141 Rue de la Cardonille, 34396 Montpellier Cedex 5, France.
Albrecht
Kenneth H
KH
Malki
Safia
S
Berta
Philippe
P
Boizet-Bonhoure
Brigitte
B
Poulat
Francis
F
D020816:Amino Acid Motifs; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D017124:Conserved Sequence; D003598:Cytoskeletal Proteins / Q000378:metabolism / Q000502:physiology*; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D009419:Nerve Tissue Proteins / Q000378:metabolism / Q000502:physiology*; D010750:Phosphoproteins; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000737:chemistry; D012708:Sertoli Cells / Q000166:cytology; D051876:Sex-Determining Region Y Protein / Q000378:metabolism*; D017923:Sodium-Hydrogen Exchangers; D013045:Species Specificity; D013347:Subcellular Fractions; D013737:Testis / Q000378:metabolism; D014162:Transfection
D003598:Cytoskeletal Proteins; C108381:GEMIN2 protein, human; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; C491561:Sip1 protein, mouse; D017923:Sodium-Hydrogen Exchangers; C497746:Sry protein, mouse; C093376:sodium-hydrogen exchanger regulatory factor; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase