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Genetic and Cytological Analysis of SPE-6, a C. elegans Tau Tubulin Kinase
Peterson, Jackson J
Peterson, Jackson J
Abstract
To acquire motility, Caenorhabditis elegans sperm must undergo the regulated cellular remodeling process of sperm activation, but in the complete absence of new protein synthesis. Nematode sperm utilize two general mechanisms to overcome this obstacle: 1) They pre-package many proteins involved in this remodeling and subsequent sperm motility within the unique Fibrous Body-Membranous Organelle (FB-MOs) complexes; 2) They employ large numbers of protein kinases and phosphatases, including SPE-6, in the acquisition and regulation of motility. Genetically, spe-6 is represented by two classes of alleles. In null alleles, spermatocytes fail to assemble FB-MOs or complete their meiotic divisions. In a second class of special alleles, spermatocytes complete their meiotic divisions, but mutant sperm activate precociously in the absence of proper extracellular signals. In this study, we contextualize SPE-6 evolutionarily by demonstrating that SPE-6 is closest in sequence homology to Tau Tubulin Kinases and that SPE-6 itself is highly conserved throughout Nematoda. We then report the first description of the dynamic sub-cellular localization pattern of SPE-6 and how this pattern changes at key steps of C. elegans sperm development. Specifically, SPE-6 localization dramatically shifts during disassembly of FB-MOs and during sperm activation. Furthermore, we characterize spe-6 sperm activation alleles at the protein and phenotypic level to determine that mis-localization or partial loss of SPE-6 expression is sufficient to induce precocious sperm activation. We further examine SPE-6 localization under a number of genetic, pharmacological, and physiological conditions and reveal that SPE-6 localization is context dependent in activated sperm. Finally, we demonstrate that SPE-6 localization changes during sperm activation correlate to a change in phosphorylation state.
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2015-05-01
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Biology