The 5'-untranslated region of the human muscle acylphosphatase mRNA has an inhibitory effect on protein expression.
The cDNA of the human muscle type acylphosphatase was isolated and characterized. The mRNA presents a very long 5'-untranslated region, covering the first half of the molecule: 175 bases of this part were cloned and prediction of the possible secondary structure showed that a very stable stem-loop structure could be formed in that region. Moreover, an additional AUG triplet was found upstream of the start codon of the protein, defining an open reading frame of 60 codons which overlapped that of acylphosphatase. The possible regulatory effect on translation of this part of the mRNA molecule was studied by means of transient transfection experiments: a 10-fold decrease in the expression of a reporter protein and a dramatic decrease in the corresponding mRNA was observed, due to the presence of the 5'-untranslated region of acylphosphatase mRNA. Mutagenesis of the upstream AUG triplet eliminated mRNA instability, leading to the hypothesis that the product of the upstream open reading frame could play a role in this mechanism.
Fiaschi
T
T
Dipartimento di Scienze Biochimiche, Università di Firenze, Florence, Italy.
Marzocchini
R
R
Raugei
G
G
Veggi
D
D
Chiarugi
P
P
Ramponi
G
G
D017766:Acid Anhydride Hydrolases / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000097024:Acylphosphatase
D017766:Acid Anhydride Hydrolases; D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D000097024:Acylphosphatase