The short 5' untranslated region of the betaA3/A1-crystallin mRNA is responsible for leaky ribosomal scanning.
Leaky ribosomal scanning allows the expression of multiple proteins from a single mRNA by occasionally skipping the first start codon, and initiating translation at a subsequent one. BetaA3- and betaA1-crystallin, two members of the beta-crystallin family of vertebrate eye lens proteins, are produced via this mechanism, of which, until now, only very few examples have been found in eukaryotic genes. Since the two start codons on the betaA3/A1 messenger lie in the same reading frame, the two translated proteins are identical, except for the 17 residues shorter N-terminal extension of betaA1-crystallin. It has been suggested that the very short leader (5-7 nucleotides) of the betaA3/A1 messenger might cause slippage at the first start codon, although the unfavorable context of this start codon might also be responsible. Using transient transfections, we now demonstrate that increasing the length of the leader sequence to 67 nucleotides indeed completely abolishes translation initiation at the second start codon, and thus expression of the betaA1-crystallin protein. Messengers having a leader of 5, 7 or 14 nucleotides all express both betaA3- and betaA1-crystallin at very similar relative levels.
Werten
P J
PJ
University of Nijmegen, Department of Biochemistry, The Netherlands.
Stege
G J
GJ
de Jong
W W
WW
D020121:5' Untranslated Regions / Q000235:genetics / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D018387:Codon, Initiator / Q000235:genetics; D006224:Cricetinae; D003459:Crystallins / Q000032:analysis / Q000235:genetics*; D004274:DNA, Recombinant; D015870:Gene Expression / Q000502:physiology; D005822:Genetic Vectors; D014176:Protein Biosynthesis / Q000502:physiology*; D020033:Protein Isoforms / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic; D020506:Untranslated Regions / Q000502:physiology*
D020121:5' Untranslated Regions; D018387:Codon, Initiator; D003459:Crystallins; D004274:DNA, Recombinant; D020033:Protein Isoforms; D012333:RNA, Messenger; D020506:Untranslated Regions