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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Active Transport, Cell Nucleus / physiology
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Adaptor Proteins, Signal Transducing / genetics
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Anaphase / physiology*
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Autophagy / drug effects
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Autophagy / physiology*
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Autophagy-Related Proteins
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Blotting, Western
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Cell Cycle Checkpoints / physiology*
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Cell Cycle Proteins / metabolism
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DNA Breaks, Double-Stranded*
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Endopeptidases / metabolism
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Green Fluorescent Proteins
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Intracellular Signaling Peptides and Proteins / metabolism*
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Nuclear Proteins / metabolism
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Protein Serine-Threonine Kinases / metabolism*
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Saccharomyces cerevisiae
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Saccharomycetales
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Securin
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Separase
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Sirolimus / pharmacology
Substances
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ATG13 protein, S cerevisiae
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Adaptor Proteins, Signal Transducing
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Autophagy-Related Proteins
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Cell Cycle Proteins
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Intracellular Signaling Peptides and Proteins
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Nuclear Proteins
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PDS1 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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Securin
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Green Fluorescent Proteins
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MEC1 protein, S cerevisiae
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Protein Serine-Threonine Kinases
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Endopeptidases
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ESP1 protein, S cerevisiae
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Separase
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Sirolimus