Actin-related protein T2 (ACTRT2) localizes to the perinuclear theca (PT) of male germ cells, yet its functional significance remains unclear. ACTRT2 is evolutionarily conserved and exhibits significant sequence similarity to other testis-specific actin-related proteins, with the highest conservation observed within the canonical actin core domain. We generated Actrt2-deficient mice which displayed male subfertility with pronounced acrosomal malformations originating during the Cap phase of acrosome biogenesis. Actrt2-deficient male mice showed reduced fertilization rate and poor blastocysts quality. Co-immunoprecipitation identified ACTRT2 interactions with PT proteins ACTRT1, ACTRT3, ACTL7A, ACTL9, PFN3, SPEM2 and CCIN while the interaction with CYLC1 was not detected. ACTRT2 overexpression in HEK293T cells altered cell morphology and F-actin distribution. Further, cytoskeletal regulator CFL1 was enriched in testis from Actrt2-deficient mice. We propose that ACTRT2 is a structural component of the PT stabilizing the acroplaxome during spermiogenesis and acrosome biogenesis by modulating actin dynamics. Finally, the high degree of sequence conservation and similarity with ACTRT1 and ACTRT3 together with their similar phenotypes when deleted, indicate that ACTRT2 shares a partial functional redundancy and compensatory capacity with other Arp proteins in testis. Taken together, these findings establish ACTRT2 as a structural regulator of sperm head architecture and male fertility in mice.
Kovacevic, A., Ordziniak, E., Hinterlang, L. D., Arevalo, L., Merges, G. E., Schneider, S., Schorle, H.
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