RNA quality control factors nucleate Clr4/SUV39H and trigger constitutive heterochromatin assembly

ABSTRACT: RNA surveillance (MTREC) and degradation (exosome) complexes nucleate Clr4/SUV39H at a heterochromatic long noncoding RNA, at which the two deacetylases, Sir2 and Clr3, also amass by distinct mechanisms. Iterative cycles of H3K9 deacetylation and methylation spread Clr4 from the nucleation center which with the help from RNAi establish heterochromatin. (this is like the abstract for the paper) To identify nuclear Sir2-interacting proteins, a protocol was adopted by which frozen intact nuclei suitable for biochemical purifications were released by low pressure, semi-automated cryogrinding of frozen yeast pellets after which the nuclei were isolated by differential centrifugation. From these nuclear extracts, FLAG-tagged Sir2 was purified and analyzed by nanoscale micro-capillary tandem mass spectrometry.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Schizosaccharomyces Pombe (ncbitaxon:4896)

SUBMITTER: Mo Motamedi   Wilhelm Haas  

PROVIDER: MSV000094160 | MassIVE | Sun Feb 25 07:50:00 GMT 2024

REPOSITORIES: MassIVE