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

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Title
ACA-specific RNA sequence recognition is acquired via the loop 2 region of MazF mRNA interferase
Author(s)
Jung Ho Park; S Yoshizumi; Y Yamaguchi; K P Wu; M Inouye
Bibliographic Citation
Proteins-Structure Function and Bioinformatics, vol. 81, no. 5, pp. 874-883
Publication Year
2013
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.
Keyword
Chimeric proteinsMazE-MazFSequence-specific endoribonucleasesTA systems
ISSN
0887-3585
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/prot.24246
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > Bio-Evaluation Center > 1. Journal Articles
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