CDC27Hs colocalizes with CDC16Hs to the centrosome and mitotic spindle and is essential for the metaphase to anaphase transition.
We have isolated cDNAs and raised antibodies corresponding to the human homologs of the S. cerevisiae CDC27 and CDC16 proteins, which are tetratrico peptide repeat (TPR)-containing proteins essential for mitosis in budding yeast. We find that the CDC27Hs and CDC16Hs proteins colocalize to the centrosome at all stages of the mammalian cell cycle, and to the mitotic spindle. Injection of affinity-purified anti-CDC27Hs antibodies into logarithmically growing HeLa cells causes a highly reproducible cell cycle arrest in metaphase with apparently normal spindle structure. We conclude that CDC27 and CDC16 are evolutionarily conserved components of the centrosome and mitotic spindle that control the onset of postmetaphase events during mitosis.
Tugendreich
S
S
Department of Molecular Biology and Genetics, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA.
Tomkiel
J
J
Earnshaw
W
W
Hieter
P
P
D000705:Anaphase / Q000187:drug effects / Q000502:physiology; D000906:Antibodies / Q000494:pharmacology; D064191:Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; D064194:Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000302:isolation & purification*; D018385:Centrosome / Q000737:chemistry*; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D008677:Metaphase / Q000187:drug effects / Q000502:physiology; D008845:Microinjections; D008969:Molecular Sequence Data; D029701:Saccharomyces cerevisiae Proteins; D017385:Sequence Homology; D008941:Spindle Apparatus / Q000737:chemistry*; D044767:Ubiquitin-Protein Ligases
D000906:Antibodies; D064191:Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; D064194:Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome; C577397:CDC16 protein, S cerevisiae; C577395:CDC16 protein, human; C577355:CDC27 protein, S cerevisiae; C577366:CDC27 protein, human; D018797:Cell Cycle Proteins; D029701:Saccharomyces cerevisiae Proteins; D044767:Ubiquitin-Protein Ligases