Effect of deletions 5' to the translation initiation sequence on the expression of an mRNA in animal cells.
To learn if an mRNA.18S rRNA interaction or a special secondary structure in the mRNA start region is essential for translation in eukaryotic cells, we constructed recombinant plasmids with the SV40 early promoter 5' to part of the Escherichia coli tufB-lacZ gene. Deletion of bases potentially complementary to the 18S rRNA highly increased the transient beta-galactosidase expressed in transfected CHO cells. Deletion of bases that fostered formation of potential hairpins with the mRNA 5'-terminus or altered the structure of the coding region reduced beta-galactosidase activity suggesting that these features of the mRNA secondary structure may be essential for initiation of translation. Computer aided analysis of the potential structure of 290 mRNAs suggests these are conserved features of the initiation region.
Ganoza
M C
MC
C.H. Best Institute, Banting & Best Department of Medical Research, University of Toronto, Ontario, Canada.
Farrow
N A
NA
An
G
G
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D003001:Cloning, Molecular; D003062:Codon; D006224:Cricetinae; D004247:DNA; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion
D003062:Codon; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D011993:Recombinant Fusion Proteins; D004247:DNA