Division of Cell and Developmental Genetics, Department of Medicine, VA Medical Center-111C5, University of California at San Francisco, San Francisco, CA 94121, USA.
Sry encodes a putative transcription factor that switches on testis differentiation during embryogenesis. Currently, the mechanism(s) by which Sry mediates such developmental process is still uncertain. To understand its gene regulation mechanism, we have utilized an in vitro affinity chromatography and proteomic strategy to identify and characterize Sry binding proteins from the mouse testis potentially involved in the formation of an Sry transcriptional complex(es). Our study has consistently identified the poly(ADP-ribose) polymerase 1 (PARP-1) as an Sry interactive protein. PARP-1 is expressed in mouse fetal gonads at the time of sex determination and co-localized with Sry in the nuclei of pre-Sertoli cells. PARP-1 could be co-immunoprecipitated with Sry in cultured cells. The interactive domains have been mapped to the HMG box of Sry and the zinc fingers of the PARP-1 protein, respectively. The Sry-PARP-1 interaction is evolutionarily conserved and it interferes with the ability of Sry in binding to its consensus sequence. In the presence of its substrate, PARP-1 poly(ADP-ribosyl)ates Sry and minimizes severely its DNA-binding activities. PARP-1 represses Sry-mediated transactivation of a reporter gene in cultured cells. Hence, PARP-1 could modulate the regulatory function(s) of Sry on its target genes in this developmental pathway.