title: DNA damage checkpoint triggers autophagy to regulate the initiation of anaphase.
abstract: Budding yeast cells suffering a single unrepaired double-strand break (DSB) trigger the Mec1 (ATR)-dependent DNA damage response that causes them to arrest before anaphase for 12-15 h. Here we find that hyperactivation of the cytoplasm-to-vacuole (CVT) autophagy pathway causes the permanent G2/M arrest of cells with a single DSB that is reflected in the nuclear exclusion of both Esp1 and Pds1. Transient relocalization of Pds1 is also seen in wild-type cells lacking vacuolar protease activity after induction of a DSB. Arrest persists even as the DNA damage-dependent phosphorylation of Rad53 diminishes. Permanent arrest can be overcome by blocking autophagy, by deleting the vacuolar protease Prb1, or by driving Esp1 into the nucleus with a SV40 nuclear localization signal. Autophagy in response to DNA damage can be induced in three different ways: by deleting the Golgi-associated retrograde protein complex (GARP), by adding rapamycin, or by overexpression of a dominant ATG13-8SA mutation.
authors.affiliation: Rosenstiel Center and Department of Biology, Brandeis University, Waltham, MA 02445, USA.
authors.forename: Farokh
authors.initials: F
authors.lastname: Dotiwala
authors.forename: Vinay V
authors.initials: VV
authors.lastname: Eapen
authors.forename: Jacob C
authors.initials: JC
authors.lastname: Harrison
authors.forename: Ayelet
authors.initials: A
authors.lastname: Arbel-Eden
authors.forename: Vikram
authors.initials: V
authors.lastname: Ranade
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yoshida
authors.forename: James E
authors.initials: JE
authors.lastname: Haber
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000705:Anaphase / Q000502:physiology*; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D000071183:Autophagy-Related Proteins; D015153:Blotting, Western; D059447:Cell Cycle Checkpoints / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D010450:Endopeptidases / Q000378:metabolism; D049452:Green Fluorescent Proteins; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D004718:Saccharomycetales; D064249:Securin; D064247:Separase; D020123:Sirolimus / Q000494:pharmacology
chemical_list: C413216:ATG13 protein, S cerevisiae; D048868:Adaptor Proteins, Signal Transducing; D000071183:Autophagy-Related Proteins; D018797:Cell Cycle Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; C578004:PDS1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D064249:Securin; D049452:Green Fluorescent Proteins; C088937:MEC1 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases; D010450:Endopeptidases; C577986:ESP1 protein, S cerevisiae; D064247:Separase; D020123:Sirolimus