Complementation of a yeast cell cycle mutant by an alfalfa cDNA encoding a protein kinase homologous to p34cdc2.
The cdc2 protein kinase plays a central role in control of the eukaryotic cell cycle of animals and yeasts. We have isolated a cDNA clone (cdc2Ms) from alfalfa (Medicago sativa L.) that is homologous to the yeast cdc2/CDC28 genes. The encoded protein is 64% identical to the yeast and mammalian counterparts and shows all the prominent structural features known from these organisms. Antibody raised against a 16-amino acid synthetic peptide with crossreactivity against p34 proteins recognized a 34-kilodalton protein in extracts of alfalfa cells. When transferred into a fission yeast, the plant cdc2 homolog can complement a temperature-sensitive cdc2 mutant. Northern analysis revealed higher transcript levels in shoots and suspension cultures than in roots. In addition to the dominant transcript of 1.4 kilobases detected in the poly(A)+fraction, 2.5- and 1.2-kilobase transcripts were detected in total RNA preparations from shoots or somatic embryos. Suspension cultures that were induced to form somatic embryos by an auxin (2,4-dichlorophenoxyacetic acid) showed fluctuations in transcription pattern during the induction period and embryogenesis.
Hirt
H
H
Institute of Microbiology and Genetics, University of Vienna, Austria.
Páy
A
A
Györgyey
J
J
Bakó
L
L
Németh
K
K
Bögre
L
L
Schweyen
R J
RJ
Heberle-Bors
E
E
Dudits
D
D
D000595:Amino Acid Sequence; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000235:genetics*; D002453:Cell Cycle; D005816:Genetic Complementation Test; D006801:Humans; D000455:Medicago sativa / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011494:Protein Kinases / Q000235:genetics*; D012568:Schizosaccharomyces / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D015345:Oligonucleotide Probes; D011494:Protein Kinases; D016203:CDC2 Protein Kinase