Skip to Content
MilliporeSigma
  • Rad9 interacts with Aft1 to facilitate genome surveillance in fragile genomic sites under non-DNA damage-inducing conditions in S. cerevisiae.

Rad9 interacts with Aft1 to facilitate genome surveillance in fragile genomic sites under non-DNA damage-inducing conditions in S. cerevisiae.

Nucleic acids research (2014-10-11)
Christos Andreadis, Christoforos Nikolaou, George S Fragiadakis, Georgia Tsiliki, Despina Alexandraki
ABSTRACT

DNA damage response and repair proteins are centrally involved in genome maintenance pathways. Yet, little is known about their functional role under non-DNA damage-inducing conditions. Here we show that Rad9 checkpoint protein, known to mediate the damage signal from upstream to downstream essential kinases, interacts with Aft1 transcription factor in the budding yeast. Aft1 regulates iron homeostasis and is also involved in genome integrity having additional iron-independent functions. Using genome-wide expression and chromatin immunoprecipitation approaches, we found Rad9 to be recruited to 16% of the yeast genes, often related to cellular growth and metabolism, while affecting the transcription of ∼2% of the coding genome in the absence of exogenously induced DNA damage. Importantly, Rad9 is recruited to fragile genomic regions (transcriptionally active, GC rich, centromeres, meiotic recombination hotspots and retrotransposons) non-randomly and in an Aft1-dependent manner. Further analyses revealed substantial genome-wide parallels between Rad9 binding patterns to the genome and major activating histone marks, such as H3K36me, H3K79me and H3K4me. Thus, our findings suggest that Rad9 functions together with Aft1 on DNA damage-prone chromatin to facilitate genome surveillance, thereby ensuring rapid and effective response to possible DNA damage events.

MATERIALS
Product Number
Brand
Product Description

Glutathione, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Supelco
Glutathione, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Phenol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Bathocuproine, sublimed grade, 99.99% trace metals basis
Sigma-Aldrich
Phenol, JIS special grade, ≥99.0%
Supelco
Phenol solution, certified reference material, 500 μg/mL in methanol
Supelco
Phenol solution, 5000 μg/mL in methanol, certified reference material
Sigma-Aldrich
Phenol, unstabilized, purified by redistillation, ≥99%
Sigma-Aldrich
Phenol, unstabilized, ReagentPlus®, ≥99%
Supelco
Bathophenanthroline, for spectrophotometric det. of Fe in serum, ≥99.0%
Sigma-Aldrich
Phenol, BioUltra, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
Phenol, ≥99%
Supelco
Phenol, PESTANAL®, analytical standard
Sigma-Aldrich
Phenol, puriss., meets analytical specification of Ph. Eur., BP, USP, ≥99.5% (GC), crystalline (detached)
Sigma-Aldrich
Phenol, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.5-100.5% (GC)
Sigma-Aldrich
Bathocuproine, 96%
Sigma-Aldrich
Phenol, ≥96.0% (calc. on dry substance, T)
Supelco
Phenol solution, 100 μg/mL in acetonitrile, PESTANAL®, analytical standard
Sigma-Aldrich
Phenol, BioUltra, for molecular biology, TE-saturated, ~73% (T)
Sigma-Aldrich
Phenol, for molecular biology
Sigma-Aldrich
L-Glutathione reduced, ≥98.0%
Sigma-Aldrich
L-Glutathione reduced, suitable for cell culture, BioReagent, ≥98.0%, powder
Sigma-Aldrich
6-Azauracil, ≥98%
Sigma-Aldrich
Phenol solution, BioReagent, Saturated with 0.01 M citrate buffer, pH 4.3 ± 0.2, for molecular biology
Sigma-Aldrich
Liquified Phenol, ≥89.0%
Sigma-Aldrich
Phenol solution, Equilibrated with 10 mM Tris HCl, pH 8.0, 1 mM EDTA, BioReagent, for molecular biology
Sigma-Aldrich
L-Glutathione reduced, BioXtra, ≥98.0%
Sigma-Aldrich
Phenol, BioXtra, ≥99.5% (GC)
Sigma-Aldrich
Phenol, natural, 97%, FG