Differential in vitro translation of the precursors of bovine pancreatic trypsin inhibitor and its isoinhibitor II is controlled by the 5'-end region of their mRNAs.
Bovine spleen inhibitor (SI II), a 58-amino-acid protein present in several bovine tissues, is an isoinhibitor of bovine pancreatic trypsin inhibitor (BPTI or aprotinin). These two proteins, which differ in seven amino-acidic residues, have very similar inhibitory activity against serine proteinases and are biosynthesized as two separate precursors of 100 residues. Higher levels of BPTI, compared to SI II, are found in bovine lung, as well as in other bovine tissues, in contrast to the level in vivo of the corresponding mRNAs. SI mRNA possesses a 90-nt 5'-end region, absent in BPTI mRNA, with an additional 5' AUG in a different open reading frame (ORF). We have used an in vitro transcription/translation system to determine the effect of this upstream region on the efficiency of SI precursor translation. Full-length SI mRNA is translated in vitro 6-fold less efficiently than BPTI mRNA. However, when SI mRNA lacks the 5' non-coding region, the translational efficiency of the 'truncated' transcript is significantly increased, reaching the same level as that of BPTI mRNA. In all cases the 10,500 Da precursor is the product of the in vitro translation. Our results indicate that the dramatic differences in translational efficiency of the mRNAs encoding BPTI and SI II in vitro parallel the different levels of the two proteins in vivo, and could be attributed to the features of the 5' non-coding region of SI mRNA.
Gambacurta
A
A
Department of Experimental Medicine and Biochemical Sciences, University of Rome Tor Vergata, Italy.
Piro
M C
MC
Ascoli
F
F
D000818:Animals; D007611:Aprotinin / Q000031:analogs & derivatives* / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D008168:Lung / Q000378:metabolism; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*
D015345:Oligonucleotide Probes; D011498:Protein Precursors; D012333:RNA, Messenger; C062659:spleen protease inhibitor II; D007611:Aprotinin