Regulation of recombination at yeast nuclear pores controls repair and triplet repeat stability.
Secondary structure-forming DNA sequences such as CAG repeats interfere with replication and repair, provoking fork stalling, chromosome fragility, and recombination. In budding yeast, we found that expanded CAG repeats are more likely than unexpanded repeats to localize to the nuclear periphery. This positioning is transient, occurs in late S phase, requires replication, and is associated with decreased subnuclear mobility of the locus. In contrast to persistent double-stranded breaks, expanded CAG repeats at the nuclear envelope associate with pores but not with the inner nuclear membrane protein Mps3. Relocation requires Nup84 and the Slx5/8 SUMO-dependent ubiquitin ligase but not Rad51, Mec1, or Tel1. Importantly, the presence of the Nup84 pore subcomplex and Slx5/8 suppresses CAG repeat fragility and instability. Repeat instability in nup84, slx5, or slx8 mutant cells arises through aberrant homologous recombination and is distinct from instability arising from the loss of ligase 4-dependent end-joining. Genetic and physical analysis of Rad52 sumoylation and binding at the CAG tract suggests that Slx5/8 targets sumoylated Rad52 for degradation at the pore to facilitate recovery from acute replication stress by promoting replication fork restart. We thereby confirmed that the relocation of damage to nuclear pores plays an important role in a naturally occurring repair process.
Department of Biology, Tufts University, Medford, Massachusetts 02155, USA;
Xiaofeng A
XA
Su
Friedrich Miescher Institute for Biomedical Research, CH-4058 Basel, Switzerland;
Vincent
V
Dion
Friedrich Miescher Institute for Biomedical Research, CH-4058 Basel, Switzerland; Faculty of Natural Sciences, University of Basel, CH-4056 Basel, Switzerland.
Susan M
SM
Gasser
Department of Biology, Tufts University, Medford, Massachusetts 02155, USA; Program in Genetics, Tufts University, Medford, Massachusetts 02155, USA; catherine.freudenreich@tufts.edu.
Catherine H
CH
Freudenreich
D004260:DNA Repair / Q000235:genetics*; D022022:Nuclear Pore / Q000378:metabolism*; D021381:Protein Transport; D011995:Recombination, Genetic; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D058207:Sumoylation; D018911:Trinucleotide Repeats / Q000235:genetics*
D029701:Saccharomyces cerevisiae Proteins
CAG repeats; Slx5/Slx8; chromosome fragility; nuclear organization; nuclear pore; repeat expansion