Cdc16p, Cdc23p and Cdc27p form a complex essential for mitosis.
Cdc16p, Cdc23p and Cdc27p are all essential proteins required for cell cycle progression through mitosis in Saccharomyces cerevisiae. All three proteins contain multiple tandemly repeated 34 amino acid tetratricopeptide repeats (TPRs). Using two independent assays, two-hybrid analysis in vivo and co-immunoprecipitation in vitro, we demonstrate that Cdc16p, Cdc23p and Cdc27p self associate and interact with one another to form a macromolecular complex. A temperature sensitive mutation in the most highly conserved TPR domain of Cdc27p results in a greatly reduced ability to interact with Cdc23p, but has no effect on interactions with wild-type Cdc27p or Cdc16p. The specificity of this effect indicates that TPRs can mediate protein-protein interactions and that this mutation may define an essential interaction for cell cycle progression in yeast. The conservation of at least two of the three proteins from yeast to man suggests that this protein complex is essential for mitosis in a wide range of eukaryotes.
Lamb
J R
JR
Department of Molecular Biology and Genetics, Johns Hopkins University, School of Medicine, Baltimore, MD 21205.
Michaud
W A
WA
Sikorski
R S
RS
Hieter
P A
PA
D000595:Amino Acid Sequence; D064173:Anaphase-Promoting Complex-Cyclosome; D000908:Antibodies, Fungal; D064191:Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; D064194:Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome; D001483:Base Sequence; D018797:Cell Cycle Proteins; D004252:DNA Mutational Analysis; D005057:Eukaryotic Cells / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005822:Genetic Vectors; D046911:Macromolecular Substances; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000254:growth & development*; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D043743:Ubiquitin-Protein Ligase Complexes; D044767:Ubiquitin-Protein Ligases; D001616:beta-Galactosidase / Q000032:analysis
D000908:Antibodies, Fungal; D064191:Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; D064194:Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome; C577397:CDC16 protein, S cerevisiae; C577355:CDC27 protein, S cerevisiae; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome; D044767:Ubiquitin-Protein Ligases; D001616:beta-Galactosidase