Identification of YAP as sperm-PAWP’s predominant binding partner in MII-arrested oocytes and the relevance of this WWI domain modular interaction to oocyte activation

PAWP, a candidate sperm-borne oocyte activating factor, induces oocyte activation and acts upstream of the calcium signalling pathway, however, PAWP’s downstream signalling pathway in oocyte cytoplasm remains to be uncovered. Data from our lab suggested that the interacting partner of PAWP, at least in the frog (Xenopus laevis) model may be YAP, a highly expressed protein in amphibian and mammalian oocytes. Therefore, the objectives of this study were to confirm that PAWP’s predominant binding partner in Xenopus laevis oocyte is YAP; to determine if mammalian oocyte activation is also dependent on PAWP-YAP interaction; and to verify that the PAWP-YAP interaction during oocyte activation is dependent on the WWI domain module. By immunohistochemistry, YAP was localized predominantly in the cytosol of metaphase II-arrested Xenopus laevis oocytes, where presumably the PAWP-YAP interaction occurs. Utilizing Far Western blotting, YAP was identified as the predominant binding partner of PAWP, in metaphase II-arrested frog (Xenopus laevis), swine (Sus scrofa) and mouse (mus musculus) oocytes. The specificity of this interaction was then tested on Far Western blotting of mouse ovarian and oocyte cytosolic extracts, by competition with both wild-type and point-mutated recombinant WWI domains derived from YAP. The removal of GST from the wild-type WWI-GST fusion protein was a requirement for effective blockage of WWI module interaction between PAWP and YAP. As expected, the mutated WWI domain was ineffective in inhibiting the PAWP-YAP interaction. To conclude, this study identified YAP as the predominant binding partner of PAWP in both amphibian and mammalian oocytes, and showed this interaction is dependent on the WWI modular interaction. The results allow us to test the functional relevance of this WWI modular interaction during oocyte activation in vivo, in the future.

The Relationship Between Coaching Behaviours and Sport Anxiety in Athletes

Previous research has identified the relationship between athlete sport anxiety and various sport outcomes (e.g., performance and dropout). For the majority of athletes involved in sport, the coach is an influential element of the competitive experience. Two hundred and twenty-eight athletes from 15 sports, completed the Sport Anxiety Scale (SAS) and the Coaching Behavior Scale for Sport (CBS-S). The predictive ability of athletes' perceived frequency of seven coaching behaviours (physical training, mental preparation, goal setting, technical skills, competition strategies, personal rapport and negative personal rapport) on four forms of sport anxiety (total anxiety, somatic anxiety, concentration disruption and worry) was examined. Results indicate that negative personal rapport was a significant predictor of all measured forms of sport anxiety while competition strategies was a significant predictor for total anxiety, concentration disruption, and worry. Other behaviours were not significant. The findings suggest that negative rapport between coach and athlete is an important contributor to athlete anxiety. In addition, behaviours that the coach demonstrates relative to competition can be influential in reducing athlete anxiety.