Nonsynonymous mutations in linker-2 of the PDR5 multidrug transporter identify a new RNA stability element

Hadiar Rahman, Andrew Rudrow, Joshua Carneglia, Sister Stephen Patrick Joly, Dante Nicotera, Michael Naldrett, John Choy, Suresh V. Ambudkar, John Golin

Research output: Contribution to journalArticle

Abstract

Analysis of synonymous mutations established that although the primary amino acid sequence remains unchanged, alterations in transcription and translation can result in significant phenotypic consequences. We report the novel observation that a series of nonsynonymous mutations in an unconserved stretch of amino acids found in the yeast multidrug efflux pump Pdr5 increases expression, thus enhancing multidrug resistance. Cycloheximide chase experiments ruled out the possibility that the increased steady-state level of Pdr5 was caused by increased protein stability. Quantitative-RT PCR experiments demonstrated that the mutants had levels of PDR5 transcript that were two to three times as high as in the isogenic wild-type strain. Further experiments employing metabolic labeling of mRNA with 4-thiouracil followed by uracil chasing showed that the half-life of PDR5 transcripts was specifically increased in these mutants. Our data demonstrate that the nucleotides encoding unconserved amino acids may be used to regulate expression and suggest that Pdr5 has a newly discovered RNA stability element within its coding region.

Original languageEnglish (US)
Pages (from-to)357-369
Number of pages13
JournalG3: Genes, Genomes, Genetics
Volume10
Issue number1
DOIs
StatePublished - Jan 1 2020

Keywords

  • ABC transporter
  • Drug resistance
  • Nonsynonymous mutation
  • Pdr5
  • mRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Fingerprint Dive into the research topics of 'Nonsynonymous mutations in linker-2 of the PDR5 multidrug transporter identify a new RNA stability element'. Together they form a unique fingerprint.

  • Cite this

    Rahman, H., Rudrow, A., Carneglia, J., Patrick Joly, S. S., Nicotera, D., Naldrett, M., Choy, J., Ambudkar, S. V., & Golin, J. (2020). Nonsynonymous mutations in linker-2 of the PDR5 multidrug transporter identify a new RNA stability element. G3: Genes, Genomes, Genetics, 10(1), 357-369. https://doi.org/10.1534/g3.119.400863