Several protein kinases have been shown to be involved in spermatogenesis. Recently, a novel subfamily of serine/threonine kinases has been isolated whose expression is limited to testis. Here, we report the fifth family member, named TSSK5, which encodes a 328 amino acid protein. RT-PCR analysis showed that TSSK5 is exclusively expressed in human testis. We isolated cAMP responsive element binding protein (CREB), a TSSK5 interacting protein via yeast two-hybrid system. The in vitro kinase assay showed that TSSK5 phosphorylated CREB at Ser-133. Using a CRE reporter system, we found that TSSK5 could stimulate the CREB/CRE responsive pathway in Hek293 cells. These results suggest that this kinase may be involved in spermatogenesis through phosphorylating CREB and then stimulating the CREB/CRE responsive pathway.