ACA-specific RNA sequence recognition is acquired via the loop 2 region of MazF mRNA interferase

Jung Ho Park, Satoshi Yoshizumi, Yoshihiro Yamaguchi, Kuen Phon Wu, Masayori Inouye

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

MazF is an mRNA interferase that cleaves mRNAs at a specific RNA sequence. MazF from E. coli (MazF-ec) cleaves RNA at A and CA. To date, a large number of MazF homologs that cleave RNA at specific three- to seven-base sequences have been identified from bacteria to archaea. MazF-ec forms a dimer, in which the interface between the two subunits is known to be the RNA substrate-binding site. Here, we investigated the role of the two loops in MazF-ec, which are closely associated with the interface of the MazF-ec dimer. We examined whether exchanging the loop regions of MazF-ec with those from other MazF homologs, such as MazF from Myxococcus xanthus (MazF-mx) and MazF from Mycobacterium tuberculosis (MazF-mt3), affects RNA cleavage specificity. We found that exchanging loop 2 of MazF-ec with loop 2 regions from either MazF-mx or MazF-mt3 created a new cleavage sequence at (A/U)(A/U)AA and C in addition to the original cleavage site, A and CA, whereas exchanging loop 1 did not alter cleavage specificity. Intriguingly, exchange of loop 2 with 8 or 12 consecutive Gly residues also resulted in a new RNA cleavage site at (A/U)(A/U)AA and C. The present study suggests a method for expanding the RNA cleavage repertoire of mRNA interferases, which is crucial for potential use in the regulation of specific gene expression and for biotechnological applications.

Original languageEnglish (US)
Pages (from-to)874-883
Number of pages10
JournalProteins: Structure, Function and Bioinformatics
Volume81
Issue number5
DOIs
StatePublished - May 2013

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biochemistry
  • Molecular Biology

Keywords

  • Chimeric proteins
  • MazE-MazF
  • Sequence-specific endoribonucleases
  • TA systems

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