UC Santa Cruz

Cell motility is essential for processes such as immune responses, wound healing, and fertilization. Caenorhabditis elegans is an excellent model for studying sperm motility, due to the worm's transparency and high sperm production. C. elegans amoeboid sperm move by extending and retracting pseudopods, which are powered internally by the dynamic polymerization and depolymerization of the nematode Major Sperm Protein (MSP). Previous biochemical studies in the parasitic nematode Ascaris suum identified MSP polymerization-activating kinase (MPAK) as a key regulator of MSP assembly within the pseudopod. Here we investigate two C. elegans homologs, Tau Tubulin Kinase 8.1 (ttbk-8.1) and Tau Tubulin Kinase 8.2 (ttbk-8.2), that arose from a C. elegans-specific gene duplication. We generated both single and double knockout mutants; single knockout animals have fertility comparable to wildtype. Self-fertilizing double knockout (ttbk-8.1/2) hermaphrodites are severely sub-fertile, and males are completely infertile. ttbk-8.1/2 males successfully transfer sperm to females; however, their sperm cannot crawl to the site of fertilization. During sperm activation, ttbk-8.1/2 sperm undergo the two key events of sperm activation (membranous organelle fusion and pseudopod extension), however the process is abnormally slow. Furthermore, the resulting pseudopods are non-motile and abnormally smooth. Localization of key proteins to the pseudopods remains unaffected. TTBK-8.1/2 colocalizes with MSP to the fibrous bodies in mature spermatocytes, distributes throughout the cytoplasm in spermatids, and is confined to pseudopods of spermatozoa. These findings suggest an essential role for TTBK-8.1/2 in timely sperm activation and pseudopod motility.