Higher order structures of the 5'-proximal region decrease the efficiency of translation of the porcine pro-opiomelanocortin mRNA.
The SP6 polymerase/promoter system was used to synthesize porcine pro-opiomelanocortin mRNAs with nucleotide sequence deletions in the 5'- as well as 3'-untranslated and coding regions. The translational efficiency of the mutant mRNAs was evaluated by cell-free translation or by monitoring the rate and extent of ribosome binding in the presence of sparsomycin. The results of these experiments indicate that specific nucleotide sequences in the 5'-untranslated and coding regions of the pro-opiomelanocortin mRNA decrease its rate of translation. Structure mapping of the mRNA with double-strand and single-strand specific nucleases suggests that these sequences can form stable secondary structures.
Chevrier
D
D
Département de Biochimie, Faculté de Médecine, Université de Montréal, Québec, Canada.
Vézina
C
C
Bastille
J
J
Linard
C
C
Sonenberg
N
N
Boileau
G
G
D000818:Animals; D001426:Bacterial Proteins; D002872:Chromosome Deletion; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D004591:Electrophoresis, Polyacrylamide Gel; D009690:Nucleic Acid Conformation; D011333:Pro-Opiomelanocortin / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D006163:Ribonuclease T1 / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D013552:Swine
D001426:Bacterial Proteins; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin; D004262:DNA Restriction Enzymes; C024404:BglII endonuclease; C024388:CTGCAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D006163:Ribonuclease T1