A dinoflagellate AAA family member rescues a conditional yeast G1/S phase cyclin mutant through increased CLB5 accumulation.
An AAA protein from the dinoflagellate Gonyaulax polyedra (GpAAA) with the unusual ability to rescue the phenotype of a yeast mutant lacking G1/S phase cyclins (cln1cln2cln3) has been isolated and the mechanism of rescue was characterized. We find that GpAAA is not a cyclin and has no similarity to any known cell cycle regulators. Instead, GpAAA forms a novel and strongly supported clade with bacterial spoIIIAA proteins and an Arabidopsis gene of unknown function. Since dinoflagellates cannot be transformed, we took advantage of the powerful genetic tools available for yeast. We find that rescue of the cln1cln2cln3 phenotype does not involve an effect on the CDK-inhibitor (CKI) Sic1p, as GpAAA does not alter the sensitivity to an inducible SIC1. Instead, Northern blot analyses show that GpAAA expression increases levels of CLB5, in agreement with the observation that GpAAA is unable to rescue the quadruple mutant cln1cln2cln3clb5. We propose that the increased transcription of CLB5 may be due to a protein remodeling function of GpAAA alleviating inhibition of the transcription factor SBF. Thus, although no known equivalents to the yeast SBF have been documented in dinoflagellates, we conclude that dinoflagellates could indeed utilize GpAAA as a cell cycle regulator.
Bertomeu
Thierry
T
Institut de Recherche en Biologie Végétale, Département de Sciences Biologiques, Université de Montréal 4101 Sherbrooke Est, Montréal, Québec, Canada H1X 2B2.
Morse
David
D
D000818:Animals; D017360:Arabidopsis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D015152:Blotting, Northern; D019926:Cyclin B / Q000235:genetics; D016213:Cyclins / Q000235:genetics*; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004141:Dinoflagellida / Q000235:genetics*; D015870:Gene Expression; D020043:Genes, Essential; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics; D015800:Protozoan Proteins / Q000235:genetics*; D012331:RNA, Fungal / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; C076558:CLB5 protein, S cerevisiae; D019926:Cyclin B; D016213:Cyclins; D016054:DNA, Protozoan; D010940:Plant Proteins; D015800:Protozoan Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins