Genetic screen for suppression of transcriptional interference reveals fission yeast 14-3-3 protein Rad24 as an antagonist of precocious Pol2 transcription termination. Academic Article uri icon

Overview

abstract

  • Expression of fission yeast Pho1 acid phosphatase is repressed under phosphate-replete conditions by transcription of an upstream prt lncRNA that interferes with the pho1 mRNA promoter. lncRNA control of pho1 mRNA synthesis is influenced by inositol pyrophosphate (IPP) kinase Asp1, deletion of which results in pho1 hyper-repression. A forward genetic screen for ADS (Asp1 Deletion Suppressor) mutations identified the 14-3-3 protein Rad24 as a governor of phosphate homeostasis. Production of full-length interfering prt lncRNA was squelched in rad24Δ cells, concomitant with increased production of pho1 mRNA and increased Pho1 activity, while shorter precociously terminated non-interfering prt transcripts persisted. Epistasis analysis showed that pho1 de-repression by rad24Δ depends on: (i) 3'-processing and transcription termination factors CPF, Pin1, and Rhn1; and (ii) Threonine-4 of the Pol2 CTD. Combining rad24Δ with the IPP pyrophosphatase-dead asp1-H397A allele caused a severe synthetic growth defect that was ameliorated by loss-of-function mutations in CPF, Pin1, and Rhn1, and by CTD phospho-site mutations T4A and Y1F. Rad24 function in repressing pho1 was effaced by mutation of its phosphate-binding pocket. Our findings instate a new role for a 14-3-3 protein as an antagonist of precocious RNA 3'-processing/termination.

publication date

  • January 25, 2022

Research

keywords

  • 14-3-3 Proteins
  • Cell Cycle Proteins
  • Gene Expression Regulation, Fungal
  • Intracellular Signaling Peptides and Proteins
  • RNA Polymerase II
  • Schizosaccharomyces
  • Schizosaccharomyces pombe Proteins
  • Transcription, Genetic

Identity

PubMed Central ID

  • PMC8789043

Scopus Document Identifier

  • 85123878766

Digital Object Identifier (DOI)

  • 10.1093/nar/gkab1263

PubMed ID

  • 34967420

Additional Document Info

volume

  • 50

issue

  • 2