The long-term objectives of the proposed research are to elucidate how calcium/calmodulin-dependent protein kinase IV (CaMKIV) participates in calcium/calmodulin (Ca2+/CaM)-mediated signal transduction cascades and regulates the cells in which it is expressed. Mice have been generated that are deficient in CaMKIV, its upstream activating kinase CaMKK beta or its alternate gene product calspermin. Camk4-/- mice are male (and female) infertile, unable to mount a type 2 immune response and exhibit profound loss of coordination and motor control. The specific defect in male infertility involves a disappearance of the protamine 2 precursor in step 15 spermatids and is similar to a defect identified in a subset of infertile human males. The immunological defects arise due to the inability to produce IL-4 in naive CD4+ T cells and, thus, are reminiscent of those identified in allergic asthma. Finally, the survival of thymocytes and cerebellar Purkinje cells is decreased in the Camk4-/- mice in a manner that may be related to transcription of genes regulated by orphan receptors of the ROR family. It is proposed to examine the molecular events in naive CD4+ T cells and thymocytes that require CaMKIV (or CaMKK beta) using transcription of IL-4 and transcription by ROR as endpoints. This will be accomplished using a combination of molecular/biochemical, functional genomic and proteomic technology. This research will define the phosphoproteome of the naive CD4+ T cell and identify the genes in thymocytes that require ROR for expression. In addition the research will determine the role for calspermin in spermiogenesis and ovulation.