title: mRNA secondary structure modulates translation of Tat-dependent formate dehydrogenase N.
abstract: Formate dehydrogenase N (FDH-N) of Escherichia coli is a membrane-bound enzyme comprising FdnG, FdnH, and FdnI subunits organized in an (alphabetagamma)3 configuration. The FdnG subunit carries a Tat-dependent signal peptide, which localizes the protein complex to the periplasmic side of the membrane. We noted that substitution of the first arginine (R5) in the twin arginine signal sequence of FdnG for a variety of other amino acids resulted in a dramatic (up to 60-fold) increase in the levels of protein synthesized. Bioinformatic analysis suggested that the mRNA specifying the first 17 codons of fdnG forms a stable stem-loop structure. A detailed mutational analysis has demonstrated the importance of this mRNA stem-loop in modulating FDH-N translation.
authors.affiliation: Department of Molecular Microbiology, John Innes Centre, Colney Lane, Norwich NR4 7UH, United Kingdom.
authors.forename: Claire
authors.initials: C
authors.lastname: Punginelli
authors.forename: Bérengère
authors.initials: B
authors.lastname: Ize
authors.forename: Nicola R
authors.initials: NR
authors.lastname: Stanley
authors.forename: Valley
authors.initials: V
authors.lastname: Stewart
authors.forename: Gary
authors.initials: G
authors.lastname: Sawers
authors.forename: Ben C
authors.initials: BC
authors.lastname: Berks
authors.forename: Tracy
authors.initials: T
authors.lastname: Palmer
mesh_terms: D019677:Artificial Gene Fusion; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis; D005560:Formate Dehydrogenases / Q000096:biosynthesis*; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D033903:Periplasmic Proteins / Q000096:biosynthesis; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D033903:Periplasmic Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D005560:Formate Dehydrogenases; D001616:beta-Galactosidase