The Dictyostelium dual-specificity kinase splA is essential for spore differentiation.
We have studied the structure and function of the Dictyostelium kinase splA. A truncated form of the splA protein exhibited primarily tyrosine kinase activity in vitro; however, it also autophosphorylated on serine and threonine residues. The kinase domain of splA exhibits approximately 38% identity to the CTR1 kinase of Arabidopsis, which is a member of the Raf family. Outside its kinase domain, splA shares homology with the byr2 kinase of S. pombe. By aligning the sequences of splA, byr2 and STE11, a homologue of byr2 in S. cerevisiae, we have identified a conserved motif that is also found in members of the Eph family of growth factor receptor tyrosine kinases. SplA is expressed throughout development with a peak during the mound stage of morphogenesis. Strains in which the splA gene had been disrupted completed fruiting body formation; however, spore cells spontaneously lysed before completing their differentiation. Northern analysis revealed the expression of the prespore marker cotB and the prestalk markers ecmA and ecmB in the mutant strain during development. The spore differentiation marker spiA was detected in the mutant spores both by northern and immunoblotting, but these cells failed to assemble spore coats. Immunoblot analysis of the developmental pattern of tyrosine phosphorylation revealed a protein that was phosphorylated in mutants but was not phosphorylated in the wild-type cells. SplA is a novel dual specificity kinase that regulates the differentiation of spore cells.
Department of Biochemistry, Stanford University School of Medicine, CA 94305, USA. gn18s@nih.gov
G H
GH
Nuckolls
N
N
Osherov
W F
WF
Loomis
J A
JA
Spudich
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000502:physiology*; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004023:Dictyostelium / Q000235:genetics* / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D008969:Molecular Sequence Data; D010641:Phenotype; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015800:Protozoan Proteins; D013172:Spores, Fungal / Q000502:physiology*; D014443:Tyrosine / Q000378:metabolism
D004271:DNA, Fungal; D016326:Extracellular Matrix Proteins; D005656:Fungal Proteins; D015800:Protozoan Proteins; C069214:ecmB protein, Dictyostelium; D014443:Tyrosine; C102073:SplA protein, Dictyostelium discoideum; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases