The WD40 propeller domain of Cdh1 functions as a destruction box receptor for APC/C substrates.
Activation of the anaphase-promoting complex/cyclosome (APC/C) by Cdc20 and Cdh1 leads to ubiquitin-dependent degradation of securin and cyclin B and thereby promotes the initiation of anaphase and exit from mitosis. Cyclin B and securin ubiquitination depend on a destruction box (D box) sequence in these proteins, but how APC/C bound to Cdc20 or Cdh1 recognizes the D box is poorly understood. By using site-specific photocrosslinking in combination with mutational analyses, we show that the D box directly interacts with an evolutionarily conserved surface on the predicted WD40 propeller structure of Cdh1 and that this interaction is essential for processive substrate ubiquitination. We further show that Cdh1 specifically crosslinks to the APC/C subunit Cdc27 and that Cdh1 binding to APC/C depends on the presence of Cdc27. Our data imply that APC/C is activated by the association of Cdh1 with Cdc27, which enables APC/C to recognize the D box of substrates via Cdh1's propeller domain.
Kraft
Claudine
C
Research Institute of Molecular Pathology, Dr. Bohr-Gasse 7, A-1030 Vienna, Austria.
Vodermaier
Hartmut C
HC
Maurer-Stroh
Sebastian
S
Eisenhaber
Frank
F
Peters
Jan-Michael
JM
D020816:Amino Acid Motifs; D064191:Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; D064199:Cdc20 Proteins; D064200:Cdh1 Proteins; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism; D019926:Cyclin B / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D064249:Securin; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
D064191:Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome; C577495:CDC20 protein, S cerevisiae; C577355:CDC27 protein, S cerevisiae; C577366:CDC27 protein, human; C577583:CDH1 protein, S cerevisiae; D064199:Cdc20 Proteins; D064200:Cdh1 Proteins; D018797:Cell Cycle Proteins; D003432:Cross-Linking Reagents; D019926:Cyclin B; D009363:Neoplasm Proteins; D010455:Peptides; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D064249:Securin; D025801:Ubiquitin; C578002:pituitary tumor-transforming protein 1, human; D044767:Ubiquitin-Protein Ligases