Characterisation of the 5'-leader sequence of tobacco mosaic virus RNA as a general enhancer of translation in vitro.
Uncapped messenger RNAs (mRNAs) encoding calf preprochymosin, chicken prelysozyme, or Escherichia coli beta-glucuronidase (GUS) were synthesized in vitro, with or without a 5'-terminal 67-nucleotide sequence (omega') derived from the untranslated 5'-leader (omega) of tobacco mosaic virus (TMV) RNA. Messenger RNAs were translated in vitro, in messenger-dependent systems derived from rabbit reticulocytes (MDL), wheat-germ (WG) or E. coli (EC). The omega' sequence enhanced expression of each mRNA in almost every translation system. While MDL was the least responsive to omega', this sequence proved particularly efficient in permitting translation of the eukaryotic mRNAs in EC, despite the absence of a consensus Shine-Dalgarno sequence in either the mRNAs or omega'. The local context of the initiation codon (AUG) in two GUS mRNA constructs did not influence the relative enhancement caused by the omega' sequence. These findings extend the utility of omega' as a general enhancer of translation for both prokaryotic and eukaryotic mRNAs in either 80S- or 70S-ribosome-based systems.
Sleat
D E
DE
Department of Virus Research, John Innes Institute, AFRC Institute of Plant Science Research, Norwich, U.K.
Gallie
D R
DR
Jefferson
R A
RA
Bevan
M W
MW
Turner
P C
PC
Wilson
T M
TM
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase; D012085:Chymosin / Q000235:genetics; D003062:Codon / Q000235:genetics; D004742:Enhancer Elements, Genetic; D004792:Enzyme Precursors / Q000235:genetics; D005814:Genes, Viral; D019868:Kanamycin Kinase; D008969:Molecular Sequence Data; D009113:Muramidase / Q000235:genetics; D010770:Phosphotransferases / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics*; D014027:Tobacco Mosaic Virus / Q000235:genetics*
D003062:Codon; D004792:Enzyme Precursors; D012333:RNA, Messenger; D012367:RNA, Viral; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; D010770:Phosphotransferases; D019868:Kanamycin Kinase; C047895:prelysozyme c; D009113:Muramidase; C034675:preprorennin; D012085:Chymosin