CDKA and CDKB kinases from Chlamydomonas reinhardtii are able to complement cdc28 temperature-sensitive mutants of Saccharomyces cerevisiae

• Published in: Protoplasma Vol. 232; no. 3-4; pp. 183 - 191
• Main Authors: Čížková, M, Pichová, A, Vítová, M, Hlavová, M, Hendrychová, J, Umysová, D, Gálová, E, Ševčovičová, A, Zachleder, V, Bišová, K
• Format: Journal Article
• Language: English
• Published: Vienna Vienna : Springer-Verlag 01.05.2008; Springer-Verlag; Springer Nature B.V
• Subjects: Algae; Amino Acid Sequence; Animals; Biomedical and Life Sciences; CDC28 homolog; CDC28 Protein Kinase, S cerevisiae - metabolism; Cell Biology; cell cycle; Chlamydomonas reinhardtii; Chlamydomonas reinhardtii - enzymology; cyclin-dependent kinase; Cyclin-dependent kinases; Cyclin-Dependent Kinases - chemistry; Cyclin-Dependent Kinases - metabolism; Genetic Complementation Test; Life Sciences; Molecular Sequence Data; Mutation - genetics; Plant Sciences; Saccharomyces cerevisiae; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - enzymology; Sequence Alignment; Temperature; Transformation, Genetic; Yeast; Zoology; Cyclin-dependent kinase; Keywords: Cell cycle; homolog; .
• ISSN: 0033-183X; 1615-6102
• DOI: 10.1007/s00709-008-0285-z

Cyclin-dependent kinases (CDK) play a key role in coordinating cell division in all eukaryotes. We investigated the capability of cyclin-dependent kinases CDKA and CDKB from the green alga Chlamydomonas reinhardtii to complement a Saccharomyces cerevisiae cdc28 temperature-sensitive mutant. The full-length coding regions of algal CDKA and CDKB cDNA were amplified by RT-PCR and cloned into the yeast expression vector pYES-DEST52, yielding pYD52-CDKA and pYD52-CDKB. The S. cerevisiae cdc28-1N strain transformed with these constructs exhibited growth at 36 °C in inducing (galactose) medium, but not in repressing (glucose) medium. Microscopic observation showed that the complemented cells had the irregular cylindrical shape typical for G₂ phase-arrested cells when grown on glucose at 36 °C, but appeared as normal budded cells when grown on galactose at 36 °C. Sequence analysis and complementation tests proved that both CDKA and CDKB are functional CDC28/cdc2 homologs in C. reinhardtii. The complementation of the mitotic phenotype of the S. cerevisiae cdc28-1N mutant suggests a mitotic role for both of the kinases.