Stability of mRNA in the hyperthermophilic archaeon Sulfolobus solfataricus

Elisabetta Bini, Vidula Dikshit, Kristi Dirksen, Melissa Drozda, Paul Blum

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

Archaea-like bacteria are prokaryotes but, in contrast, use eukaryotic-like systems for key aspects of DNA, RNA, and protein metabolism. mRNA is typically unstable in bacteria and stable in eukaryotes, but little information is available about mRNA half-lives in archaea. Because archaea are generally insensitive to antibiotics, examination of mRNA stability in the hyperthermophile, Sulfolobus solfataricus, required the identification of transcription inhibitors for half-life determinations. An improved lacs promoter-dependent in vitro transcription system was used to assess inhibitor action. Efficient inhibitors were distinguished as blocking both lacSp transcription in vitro and the incorporation of 3H-uracil into bulk RNA in vivo. Actinomycin D was the most stable and potent compound identified. A survey of transcript chemical half-lives normalized to levels of the signal recognition particle 7S RNA ranged from at least 2 h for tfb1, a transcription factor TFIIB paralog, to a minimum of 6.3 min for gln1, one of three glutamine synthetase paralogs. Transcript half-lives for other mRNAs were: 2 h, superoxide dismutase (sod); 37.5 min, glucose dehydrogenase (dhg1); 25 min, alpha-glucosidase (malA); and 13.5 min, transcription factor TFIIB-2 (tfb2) resulting in a minimum average half-life of 54 min. These are the first mRNA half-lives reported for a hyperthermophile or member of the crenarchaea. The unexpected stability of several transcripts has important implications for gene expression and mRNA degradation in this organism.

Original languageEnglish (US)
Pages (from-to)1129-1136
Number of pages8
JournalRNA
Volume8
Issue number9
DOIs
Publication statusPublished - Sep 1 2002

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Keywords

  • Archaea
  • Hyperthermophiles
  • Transcription inhibitors
  • mRNA decay

ASJC Scopus subject areas

  • Molecular Biology

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