Cdc6 is an unstable protein whose de novo synthesis in G1 is important for the onset of S phase and for preventing a 'reductional' anaphase in the budding yeast Saccharomyces cerevisiae.
S phase entry depends on cyclin-dependent kinases whose activation during late G1 due partly to the synthesis of unstable cyclin subunits. We identify here a second type of unstable protein, Cdc6, whose synthesis during G1 is important for initiation of DNA replication. The CDC6 gene is normally transcribed at the end of mitosis, but in cells with a prolonged G1 phase there is a second burst of transcription in late G1. The former is due to Swi5, while the latter is due to MBF or SBF transcription factors. Small G1 cells that cannot synthesize Cdc6 in late G1 progress through S phase very slowly. Cells that transcribe CDC6 neither at the end of mitosis nor in late G1 fail to replicate DNA but, despite this, undergo mitosis and produce daughter cells with fractional DNA contents. This 'reductional' anaphase occurs with almost wild-type kinetics and depends on the activity of G2 cyclins. Thus, cells that fail to duplicate chromosomes due to a cdc6 defect cannot prevent the onset of mitosis, unlike other mutants with replication defects. We show, by fluorescence in situ hybridization, that chromosomes which remain unduplicated due to a lack of Cdc6 synthesis are segregated intact to spindle poles during the 'reductional' anaphase.
Piatti
S
S
Research Institute of Molecular Pathology, Vienna, Austria.
Lengauer
C
C
Nasmyth
K
K
D000705:Anaphase / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000096:biosynthesis* / Q000502:physiology; D004261:DNA Replication; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000378:metabolism; D016193:G1 Phase; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D006238:Haploidy; D008715:Methionine / Q000502:physiology; D008938:Mitosis; D008957:Models, Genetic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012331:RNA, Fungal / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000254:growth & development*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000378:metabolism; D015533:Transcriptional Activation
C066503:CDC6 protein, S cerevisiae; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C083209:MBP1 protein, S cerevisiae; D012331:RNA, Fungal; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C081540:SBF protein, S cerevisiae; C080218:SWI5 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D008715:Methionine