Isolation and characterization of a Paramecium cDNA clone encoding a putative serine/threonine protein kinase.
Mitogen-activated protein (MAP) kinases, a closely related family of protein kinases, are involved in cell cycle regulation and differentiation in yeast and human cells. They have not been documented in ciliates. We used PCR to amplify DNA sequences of a ciliated protozoan--Paramecium caudatum--using primers corresponding to amino acid sequences that are common to MAP kinases. We isolated and sequenced one putative MAP kinase-like serine/threonine kinase cDNA from P. caudatum. This cDNA, called pcstk1 (Paramecium caudatum Serine/Threonine Kinase 1) shared approximately 35% amino acid identity with MAP kinases from yeast. MAP kinases are activated by phosphorylation of specific threonine and tyrosine residues. These two amino acid residues are conserved in the PCSTK1 sequence at positions Thr 159 and Tyr 161. The PSTAIRE motif, which is characteristic of the CDK2 gene family, cannot be found in ORF of PCSTK1. The highest homology score was to human STK9, which contains MAP type kinase domains. Comparisons of expression level have shown that pcstk1 is expressed equally in cells at different stages (sexual and asexual). We discussed the possibility, as in other organisms, that a family of MAP kinase genes exists in P. caudatum.
Wada
Satoru
S
Graduate School of Life Sciences, Tohoku University, Aoba Aramaki, Sendai 980-8578, Japan. wada@saitama-med.ac.jp
Watanabe
Tsuyoshi
T
D020816:Amino Acid Motifs; D000818:Animals; D002999:Clone Cells / Q000201:enzymology; D018076:DNA, Complementary; D016054:DNA, Protozoan / Q000235:genetics*; D006801:Humans; D008018:Life Cycle Stages; D010247:Paramecium / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D018076:DNA, Complementary; D016054:DNA, Protozoan; D017346:Protein Serine-Threonine Kinases; C114482:CDKL5 protein, human