title: The role of the loop 1 region in MazFbs mRNA interferase from Bacillus subtilis in recognition of the 3' end of the RNA substrate.
abstract: MazFbs is an mRNA interferase from Bacillus subtilis specifically recognizing UACAU. The X-ray structure of its complex with an RNA substrate has been also solved. When its amino acid sequence is compared with that of MazFhw, an mRNA interferase from a highly halophilic archaeon, recognizing UUACUCA, the 9-residue loop-1 region is highly homologous except that the V16V17sequence in MazFbs is replaced with TK in MazFhw. Thus, we examined the role of the VV sequence in RNA substrate recognition by replacing it with TK, GG, AA or LL. The substitution mutants thus constructed showed significant differences in cleavage specificity using MS2 phage RNA. The primer extension analysis of the cleavage sites revealed that the VV sequence plays an important role in the recognition of the 3'-end base of the RNA substrate.
authors.affiliation: Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway, NJ 08854, USA.
authors.forename: Yojiro
authors.initials: Y
authors.lastname: Ishida
authors.affiliation: Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway, NJ 08854, USA.
authors.forename: Keiko
authors.initials: K
authors.lastname: Inouye
authors.affiliation: Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway, NJ 08854, USA. Electronic address: inouye@cabm.rutgers.edu.
authors.forename: Masayori
authors.initials: M
authors.lastname: Inouye
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004722:Endoribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D017909:Levivirus / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D012333:RNA, Messenger; D004722:Endoribonucleases
keywords: Bacillus subtilis; MS2 phage RNA; MazF; Site-directed mutagenesis