mRNA secondary structure modulates translation of Tat-dependent formate dehydrogenase N.
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.
Punginelli
Claire
C
Department of Molecular Microbiology, John Innes Centre, Colney Lane, Norwich NR4 7UH, United Kingdom.
Ize
Bérengère
B
Stanley
Nicola R
NR
Stewart
Valley
V
Sawers
Gary
G
Berks
Ben C
BC
Palmer
Tracy
T
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
D029968:Escherichia coli Proteins; D033903:Periplasmic Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D005560:Formate Dehydrogenases; D001616:beta-Galactosidase