The cdk5 homologue, crp, regulates endocytosis and secretion in dictyostelium and is necessary for optimum growth and differentiation.
Dictyostelium Crp is a member of the cyclin-dependent kinase (Cdk) family of proteins. It is most related in sequence to mammalian Cdk5, which unlike other members of the family, has functions that are unrelated to the cell cycle. In order to better understand the function of Crp in Dictyostelium, we overexpressed a dominant negative form, Crp-D144N, under the control of the actin 15 promoter. Cells overexpressing Crp-D144N exhibit a reduced growth rate in suspension culture and reduced rates of fluid-phase endocytosis and phagocytosis. There is no reduction in Cdc2 kinase activity in extracts from cells overexpressing Crp-D144N, suggesting that the growth defect is not due to inhibition of Cdc2. In addition to the growth defect, the act15::crp-D144N transformants aggregate at a slower rate than wild-type cells and form large aggregation streams. These eventually break up to form small aggregates and most of these do not produce mature fruiting bodies. The aggregation defect is fully reversed in the presence of wild-type cells but terminal differentiation is only partially rescued. In act15::crp-D144N transformants, the countin component of the counting factor, a secreted protein complex that regulates the breakup of streams, mostly appears outside the cell as degradation products and the reduced level of the intact protein may at least partially account for the initial formation of the large aggregation streams. Our observations indicate that Crp is important for both endocytosis and efflux and that defects in these functions lead to reduced growth and aberrant development.
Sharma
Shiv K
SK
Department of Microbiology and Immunology, The University of British Columbia, Vancouver, Canada.
Brock
Debra A
DA
Ammann
Robin R
RR
DeShazo
Tiffany
T
Khosla
Meenal
M
Gomer
Richard H
RH
Weeks
Gerald
G
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D051360:Cyclin-Dependent Kinase 5; D018844:Cyclin-Dependent Kinases / Q000235:genetics; D004023:Dictyostelium / Q000166:cytology / Q000502:physiology*; D004705:Endocytosis / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D015800:Protozoan Proteins / Q000378:metabolism; D015398:Signal Transduction
D015800:Protozoan Proteins; C084139:Crp protein, Dictyostelium; D051360:Cyclin-Dependent Kinase 5; D017346:Protein Serine-Threonine Kinases; D018844:Cyclin-Dependent Kinases