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PLoS Genet. 2014 May 29;10(5):e1004334. doi: 10.1371/journal.pgen.1004334. eCollection 2014.

The proper splicing of RNAi factors is critical for pericentric heterochromatin assembly in fission yeast.

Author information

  • 1Department of Biological Sciences, Columbia University, New York, New York, United States of America.
  • 2Department of Pathology and Cell Biology, Columbia University College of Physicians and Surgeons, New York, New York, United States of America.
  • 3Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, United States of America.

Abstract

Heterochromatin preferentially assembles at repetitive DNA elements, playing roles in transcriptional silencing, recombination suppression, and chromosome segregation. The RNAi machinery is required for heterochromatin assembly in a diverse range of organisms. In fission yeast, RNA splicing factors are also required for pericentric heterochromatin assembly, and a prevailing model is that splicing factors provide a platform for siRNA generation independently of their splicing activity. Here, by screening the fission yeast deletion library, we discovered four novel splicing factors that are required for pericentric heterochromatin assembly. Sequencing total cellular RNAs from the strongest of these mutants, cwf14Δ, showed intron retention in mRNAs of several RNAi factors. Moreover, introducing cDNA versions of RNAi factors significantly restored pericentric heterochromatin in splicing mutants. We also found that mutations of splicing factors resulted in defective telomeric heterochromatin assembly and mis-splicing the mRNA of shelterin component Tpz1, and that replacement of tpz1+ with its cDNA partially rescued heterochromatin defects at telomeres in splicing mutants. Thus, proper splicing of RNAi and shelterin factors contributes to heterochromatin assembly at pericentric regions and telomeres.

PMID:
24874881
PMCID:
PMC4038458
DOI:
10.1371/journal.pgen.1004334
[PubMed - indexed for MEDLINE]
Free PMC Article
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