A novel Dictyostelium Cdk8 is required for aggregation, but is dispensable for growth.
When Dictyostelium cells starve, they express genes necessary for aggregation. Using insertional mutagenesis, we have isolated a mutant that does not aggregate upon starvation and that forms small plaques on bacterial lawns, thus indicating slow growth. Sequencing of the mutated locus showed a strong similarity to the catalytic domain of cdc2-related kinase genes. Phylogenetic analysis further indicated that the amino acid sequence was more close to cyclin-dependent kinase 8 than to the sequence of other cyclin-dependent kinases. Thus, we designated this gene as Ddcdk8. The Ddcdk8-null cells do not aggregate and grow somewhat more slowly than parental cells when being shaken in axenic medium or laid on bacterial plates. To confirm whether these defective phenotypes were caused by disruption of this gene, the Ddcdk8-null cells were complemented with DdCdk8 protein expressed from an endogenous promoter, but not an actin promoter, and when the complemented cells were then allowed to grow on a bacterial lawn, they began to aggregate as the food supply was depleted and finally became fruiting bodies. The results suggest that properly regulated DdCdk8 activity is essential for aggregation. Because, when starved, Ddcdk8-null cells do not express the acaA transcripts required for aggregation, we deduce that Ddcdk8 is epistatic for acaA expression, indicating that the DdCdk8 products may regulate expression of acaA and/or other genes.
Takeda
Kosuke
K
Division of Biological Sciences, Graduate School of Science, Hokkaido University Sapporo, Japan.
Saito
Tamao
T
Ochiai
Hiroshi
H
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D004023:Dictyostelium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D008969:Molecular Sequence Data; D010802:Phylogeny; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D017422:Sequence Analysis, DNA
D015800:Protozoan Proteins; D018844:Cyclin-Dependent Kinases