Differential phosphorylation of RNA polymerase III and the initiation factor TFIIIB in saccharomyces cerevisiae

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Abstract

The production of ribosomes and tRNAs for protein synthesis has a high energetic cost and is under tight transcriptional control to ensure that the level of RNA synthesis is balanced with nutrient availability and the prevailing environmental conditions. In the RNA polymerase (pol) III system in yeast, nutrients and stress affect transcription through a bifurcated signaling pathway in which protein kinase A (PKA) and TORC1 activity directly or indirectly, through downstream kinases, alter the phosphorylation state and function of the Maf1 repressor and Rpc53, a TFIIF-like subunit of the polymerase. However, numerous lines of evidence suggest greater complexity in the regulatory network including the phosphoregulation of other pol III components. To address this issue, we systematically examined all 17 subunits of pol III along with the three subunits of the initiation factor TFIIIB for evidence of differential phosphorylation in response to inhibition of TORC1. A relatively high stoichiometry of phosphorylation was observed for several of these proteins and the Rpc82 subunit of the polymerase and the Bdp1 subunit of TFIIIB were found to be differentially phosphorylated. Bdp1 is phosphorylated on four major sites during exponential growth and the protein is variably dephosphorylated under conditions that inhibit tRNA gene transcription. PKA, the TORC1-regulated kinase Sch9 and protein kinase CK2 are all implicated in the phosphorylation of Bdp1. Alanine substitutions at the four phosphosites cause hyper-repression of transcription indicating that phosphorylation of Bdp1 opposes Maf1-mediated repression. The new findings suggest an integrated regulatory model for signaling events controlling pol III transcription.

Original languageEnglish (US)
Article numbere0127225
JournalPLoS One
Volume10
Issue number5
DOIs
StatePublished - May 13 2015

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Transcription Factor TFIIIB
RNA Polymerase III
Peptide Initiation Factors
Phosphorylation
DNA-directed RNA polymerase
Yeast
Saccharomyces cerevisiae
Transcription
phosphorylation
transcription (genetics)
cAMP-dependent protein kinase
Transfer RNA
Cyclic AMP-Dependent Protein Kinases
Nutrients
phosphotransferases (kinases)
Phosphotransferases
Casein Kinase II
Food
Protein Subunits
stoichiometry

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Differential phosphorylation of RNA polymerase III and the initiation factor TFIIIB in saccharomyces cerevisiae. / Lee, Jaehoon; Moir, Robyn D.; Willis, Ian M.

In: PLoS One, Vol. 10, No. 5, e0127225, 13.05.2015.

Research output: Contribution to journalArticle

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