Molecular mechanisms of ribosomal protein gene coregulation

Rohit Reja, Vinesh Vinayachandran, Sujana Ghosh, B. Franklin Pugh

Research output: Contribution to journalArticlepeer-review

52 Scopus citations

Abstract

The 137 ribosomal protein genes (RPGs) of Saccharomyces provide a model for gene coregulation. We examined the positional and functional organization of their regulators (Rap1 [repressor activator protein 1], Fhl1, Ifh1, Sfp1, and Hmo1), the transcription machinery (TFIIB, TFIID, and RNA polymerase II), and chromatin at near-base-pair resolution using ChIP-exo, as RPGs are coordinately reprogrammed. Where Hmo1 is enriched, Fhl1, Ifh1, Sfp1, and Hmo1 cross-linked broadly to promoter DNA in an RPG-specific manner and demarcated by general minor groove widening. Importantly, Hmo1 extended 20–50 base pairs (bp) downstream from Fhl1. Upon RPG repression, Fhl1 remained in place. Hmo1 dissociated, which was coupled to an upstream shift of the +1 nucleosome, as reflected by the Hmo1 extension and core promoter region. Fhl1 and Hmo1 may create two regulatable and positionally distinct barriers, against which chromatin remodelers position the +1 nucleosome into either an activating or a repressive state. Consistent with in vitro studies, we found that specific TFIID subunits, in addition to cross-linking at the core promoter, made precise cross-links at Rap1 sites, which we interpret to reflect native Rap1–TFIID interactions. Our findings suggest how sequence-specific DNA binding regulates nucleosome positioning and transcription complex assembly >300 bp away and how coregulation coevolved with coding sequences.

Original languageEnglish (US)
Pages (from-to)1942-1954
Number of pages13
JournalGenes and Development
Volume29
Issue number18
DOIs
StatePublished - Sep 15 2015
Externally publishedYes

Keywords

  • ChIP-seq
  • Chromatin
  • Gene regulation mechanisms
  • Transcription factor

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

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