title: The anaphase promoting complex regulates yeast lifespan and rDNA stability by targeting Fob1 for degradation.
abstract: Genomic stability, stress response, and nutrient signaling all play critical, evolutionarily conserved roles in lifespan determination. However, the molecular mechanisms coordinating these processes with longevity remain unresolved. Here we investigate the involvement of the yeast anaphase promoting complex (APC) in longevity. The APC governs passage through M and G1 via ubiquitin-dependent targeting of substrate proteins and is associated with cancer and premature aging when defective. Our two-hybrid screen utilizing Apc5 as bait recovered the lifespan determinant Fob1 as prey. Fob1 is unstable specifically in G1, cycles throughout the cell cycle in a manner similar to Clb2 (an APC target), and is stabilized in APC (apc5(CA)) and proteasome (rpn10) mutants. Deletion of FOB1 increased replicative lifespan (RLS) in wild type (WT), apc5(CA), and apc10 cells, and suppressed apc5(CA) cell cycle progression and rDNA recombination defects. Alternatively, increased FOB1 expression decreased RLS in WT cells, but did not reduce the already short apc5(CA) RLS, suggesting an epistatic interaction between apc5(CA) and fob1. Mutation to a putative L-Box (Fob1(E420V)), a Destruction Box-like motif, abolished Fob1 modifications, stabilized the protein, and increased rDNA recombination. Our work provides a mechanistic role played by the APC to promote replicative longevity and genomic stability in yeast.
authors.affiliation: Department of Anatomy and Cell Biology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5E5, Canada.
authors.forename: Johannes
authors.initials: J
authors.lastname: Menzel
authors.forename: Mackenzie E
authors.initials: ME
authors.lastname: Malo
authors.forename: Cynthia
authors.initials: C
authors.lastname: Chan
authors.forename: Martin
authors.initials: M
authors.lastname: Prusinkiewicz
authors.forename: Terra G
authors.initials: TG
authors.lastname: Arnason
authors.forename: Troy A A
authors.initials: TA
authors.lastname: Harkness
mesh_terms: D064173:Anaphase-Promoting Complex-Cyclosome / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002453:Cell Cycle; D004271:DNA, Fungal / Q000378:metabolism*; D004275:DNA, Ribosomal / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D042822:Genomic Instability; D009154:Mutation; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000254:growth & development* / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal; D005656:Fungal Proteins; D064173:Anaphase-Promoting Complex-Cyclosome
keywords: Fob1; Saccharomyces cerevisiae; anaphase promoting complex; replicative lifespan; two-hybrid screen