TMEM225: a possible protein phosphatase 1γ2 (PP1γ2) regulator localizes to the equatorial segment in mouse spermatozoa.

Tmem225 encodes a putative four-transmembrane domain protein that has an RVxF motif, which is known to be a consensus site for interacting with serine/threonine protein phosphatase 1 (PP1). We previously identified Tmem225 as one of 53 spermatogenesis-associated transmembrane protein genes, with no known physiological function. In this study, we investigated the expression and molecular characteristics of TMEM225 in mice. Tmem225 production was found to be specific to testicular germ cells, with expression increasing during spermatogenesis. In mature spermatozoa, TMEM225 is localized to the equatorial segment of the acrosome but not to the midpiece or tail. TMEM225 appears to be an outer and/or inner acrosomal membrane protein that is lost from the dorsal region of the acrosome after the acrosome reaction. TMEM225 interacts with PP1 in vivo, and a pull-down assay revealed that the carboxy-terminal region of TMEM225 can bind to PP1γ2, the predominant isoform of PP1 in male germ cells. In addition, TMEM225 inhibited PP1γ2 activity in vitro via its RVxF motif. Our results suggest that in mice, TMEM225 is involved in the differentiation and function of spermatozoa through the regulation of PP1γ2 activity, which is necessary for normal spermatogenesis as well as spermatozoa capacitation and motility.