The C-Box Region of MAF1 Regulates Transcriptional Activity and Protein Stability

J Mol Biol. 2017 Jan 20;429(2):192-207. doi: 10.1016/j.jmb.2016.12.012. Epub 2016 Dec 13.

Abstract

MAF1 is a conserved negative regulator of RNA polymerase (pol) III and intracellular lipid homeostasis across species. Here, we show that the MAF1 C-box region negatively regulates its activity. Mutations in Caenorhabditis elegans mafr-1 that truncate the C-box retain the ability to inhibit the transcription of RNA pol III targets, reduce lipid biogenesis, and lower reproductive output. In human cells, C-box deletion of MAF1 leads to increased MAF1 nuclear localization and enhanced repression of ACC1 and FASN, but with impaired repression of RNA pol III targets. Surprisingly, C-box mutations render MAF1 insensitive to rapamycin, further defining a regulatory role for this region. Two MAF1 species, MAF1L and MAF1S, are regulated by the C-box YSY motif, which, when mutated, alters species stoichiometry and proteasome-dependent turnover of nuclear MAF1. Our results reveal a role for the C-box region as a critical determinant of MAF1 stability, activity, and response to cellular stress.

Keywords: Caenorhabditis elegans; MAF1; RNA polymerase III; lipid metabolism; mTOR.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism
  • Gene Expression Regulation
  • HEK293 Cells
  • Hep G2 Cells
  • Homeostasis
  • Humans
  • Protein Stability*
  • RNA Polymerase III / metabolism*
  • Repressor Proteins / genetics*
  • Repressor Proteins / metabolism
  • Sirolimus / pharmacology
  • Transcriptional Activation*

Substances

  • Caenorhabditis elegans Proteins
  • MAFR-1 protein, C elegans
  • Repressor Proteins
  • RNA Polymerase III
  • Sirolimus