title: ACA-specific RNA sequence recognition is acquired via the loop 2 region of MazF mRNA interferase.
abstract: MazF is an mRNA interferase that cleaves mRNAs at a specific RNA sequence. MazF from E. coli (MazF-ec) cleaves RNA at A^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^C in addition to the original cleavage site, A^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^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.
authors.affiliation: Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongwon-gun, Chungcheongbuk-do, Republic of Korea.
authors.forename: Jung-Ho
authors.initials: JH
authors.lastname: Park
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yoshizumi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: Kuen-Phon
authors.initials: KP
authors.lastname: Wu
authors.forename: Masayori
authors.initials: M
authors.lastname: Inouye
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004722:Endoribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007218:Industrial Microbiology; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D016941:Myxococcus xanthus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C422955:MazF protein, E coli; D010455:Peptides; D012329:RNA, Bacterial; D012333:RNA, Messenger; C011080:polyglycine; D004722:Endoribonucleases