Mg2+ facilitates leader peptide translation to induce riboswitch-mediated transcription termination

Guang Zhao, Wei Kong, Natasha Weatherspoon-Griffin, Josephine Clark-Curtiss, Yixin Shi

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

We have characterized a 17-residue peptide, MgtL, which is translated specifically in high Mg2+ from an open reading frame (ORF) embedded in the Mg2+ riboswitch domain, previously identified in the 5′ leader region of Mg2+ transporter gene mgtA in Salmonella. We demonstrate that mgtL translation is required to prematurely terminate mgtA transcription. Abrogation of mgtL translation by mutation of its start codon results in transcription of the mgtA-coding region in high Mg2+, suggesting that ribosome stalling is not required for preventing premature transcription termination. Consistently, the Mg2+ riboswitch responds to cytoplasmic Mg2+, but not to proline or arginine, both repeatedly present in the MgtL sequence, to mediate mgtL translation-coupled regulation. RNA structural probing and nucleotide substitution analysis show that the riboswitch loop A region alters base pairing in response to Mg2+, and favours stem-loop A1 in high Mg2+, subsequently opening the ribosome-binding sequence for mgtL translation. Presumably, mgtL ORF directs translation to localize a ribosome in cis to act on downstream RNA in a manner similar to some upstream ORFs in prokaryotes and eukaryotes.

Original languageEnglish (US)
Pages (from-to)1485-1496
Number of pages12
JournalEMBO Journal
Volume30
Issue number8
DOIs
StatePublished - Apr 20 2011

Keywords

  • Mg riboswitch
  • premature termination of transcription
  • the 50 leader region
  • the leader peptide

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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