Protein kinase D (PKD, also called PKCmicro) was identified as a. serine/threonine kinase with potentially important roles in growth factor. as well as in stress-induced signaling. Moreover, PKD has emerged is an. important regulator of plasma membrane enzymes and receptors, mediating. cross-talk between different signaling systems, with functions in processes. as diverse as cell proliferation, apoptosis, immune cell regulation, tumor. cell invasion, survival, and regulation of Golgi vesicle fission and. organization. PKD isoforms are protein kinase C (PKC) effectors in. diacylglycerol (DAG)-regulated signaling pathways. Understanding the roles. of PKDs in homeostasis and signal transduction requires characterization of. functions controlled by PKDs in vivo. Caenorhabditis elegans and mammals. share conserved signaling mechanisms, molecules, and pathways. Thus,. characterization of the C.elegans PKDs and their substrates could yield. insights into regulation and functions that apply to all eukaryotic PKDs.. Kidins220, Kinase D interacting substrate of 220 kDa, also known as ankyrin. repeat-rich membrane spanning, or ARMS, is a novel integral membrane. protein abundantly expressed in the brain in mammals, the first known. physiological substrate of PKD, and encoded by a single and highly. conserved gene in C.elegans, Drosophila melanogaster, and mammals. We have. shown that Kidins220 is present in lipid rafts in brain and cells of. nervous origin, and that PKD translocates to these membrane microdomains. after phorbol ester stimulation. C.elegans Kidins220 (
tag-144, F36H1.2). shows 77,3% sequence homology with the rat protein, suggesting great. functional significance. F36H1.2 is expressed in several tissues in the. worm, including intestine, excretory cell, body wall muscle, and various. neurons. By computational sequence comparison, two isoforms of PKD have. been identified in C.elegans, T25e12.4 and W09c5.5.. We obtained the transgenic strains consisting of C.elegans Kidins220, and. both PKDs promoter::GFP fusion constructs for the in vivo analysis of the. expression of these genes in C.elegans, and we are studying them by DIC and. fluorescence microscopy. Also, we have constructed our own transgenic. strains expressing the entire promoter/gene/3''-UTR::GFP, have raised. antibodies against the three proteins, and are analysing them, We performed. the taste adaptation assay on the C.elegans Kidins220 null animal,. observing a mutant behavior, although its salt attraction behavior was wild. type. Also, Kidins220 null animals show a phenotype of slow development,. consistant with the RNAi data. We visualized the embryo development by 4D. microscopy and detected a series of abnormalities since very early, such as. multinucleated cells. When staining with anti-tubulin antibodies and DNA. stain, we observed another problems, in synchronicity of the divisions and. the axis of polarization of the spindles, cells without DNA, and with extra. DNA, etc, that we are investigating. Do not add objects such as pictures, boxes, headers, footers, footnotes,. etc.