Nuclear localization of the testis determining gene product SRY.

Abstract

Abstract. We have studied the expression of the human SRY protein (termed p27SRY) in two different cell lines by using specific antibodies. Confocal microscopy enabled us to localize p27SRY precisely in the nucleus in a discrete punctuate pattern. Furthermore, through microinjection experiments, we have demonstrated that the localization of the p27SRY protein into the nucleus was an event involving the NH2-terminal part of the high mobility group (HMG) domain. With the help of several synthetic peptides and various p27SRY mutants, we have characterized a bipartite basic motif in this part of the protein corresponding to a nuclear localization signal. This nuclear localization signal appears to be highly conserved in SRY box-and HMB box-containing proteins, suggesting common properties of nuclear targeting within the HMG box protein family.

The Human Testis Determining Factor SRY Binds a Nuclear Factor Containing PDZ Protein Interaction Domains

“…2A, this antibody detects the expected 34-kDa protein in nuclear extracts and insoluble nuclear fractions (i.e. chromatin and nucleoskeleton) derived from a human NT2D1 cell line, a pluripotent embryonic cell line positive for SRY expression (4,20). In coprecipitation assays, in vitro translated SRY protein (expected size of 27 kDa) associated specifically with SIP-1 fused to GST but not GST alone or with glutathione-Sepharose beads (Fig.…”

“…SRY encodes a small nuclear protein of 204 residues comprising three distinct domains, with a central domain of about 78 amino acids called the high mobility group (HMG) 1 box. This central domain includes a nuclear localization signal (4), and in vitro studies of the human SRY protein have demonstrated its sequence-specific DNA binding through this HMG box (5). To date, no function has been ascribed to the regions of the human SRY protein outside the HMG box, whereas in mouse Sry, the Cterminal part of the protein can function as a transcriptional activator (6).…”

A study of goat SRY protein expression suggests putative new roles for this gene in the developing testis of a species with long‐lasting SRY expression

“…between 25 and 36 dpc, gonads were not separated from mesonephroi. Wilhelm et al, 2005), or in human cells and tissues (Poulat et al, 1995;Salas-Cortés et al, 1999).…”

“…Until now, SRY antibodies were only available against mouse and human proteins (Wilhelm et al, 2005;Bradford et al, 2007;Poulat et al, 1995;Salas-Cortés et al, 1999). In humans, the first SRY antibody was polyclonal and produced against a 17 amino-acid peptide located in the Nterminal part of the protein (residues 40 to 56).…”

“…This antibody has been used to study the sub-cellular localization of SRY, and indeed, the protein has been detected in the nucleus of both human cell lines such as NT2D1, and 8-week embryonic XY gonads. In both situations, nuclear SRY staining revealed a punctuate pattern (Poulat et al, 1995). Furthermore, a monoclonal antibody against human SRY was produced and used on XY gonads at four developmental stages ($6 weeks of gestation, 26-week-old fetus, 1-month-old infant and 32-year-old adult).…”