Molecular mechanism of polyamine stimulation of the synthesis of oligopeptide-binding protein
- Resource Type
- Authors
- Masato Yuguchi; Tomoko Saisho; Kazuei Igarashi; Keiko Kashiwagi
- Source
- The Journal of biological chemistry. 272(7)
- Subject
- Untranslated region
Transcription, Genetic
Lipoproteins
Molecular Sequence Data
Biology
Biochemistry
chemistry.chemical_compound
Bacterial Proteins
Transcription (biology)
Protein biosynthesis
Polyamines
RNA, Messenger
Molecular Biology
Messenger RNA
Base Sequence
RNA
Cell Biology
Molecular biology
Open reading frame
chemistry
Protein Biosynthesis
Mutagenesis, Site-Directed
Nucleic Acid Conformation
Polyamine
Carrier Proteins
Oligopeptide binding
- Language
- ISSN
- 0021-9258
Polyamine stimulation of the synthesis of oligopeptide-binding protein (OppA) was shown to occur mainly at the level of translation by measuring OppA synthesis and its mRNA level. Several artificial oppA genes were constructed by site-directed mutagenesis. These synthesize different kinds of OppA mRNAs: mRNAs differing in the size of 5'-untranslated region; mRNAs having the Shine-Dalgarno (SD) sequence in a different position; mRNAs having different secondary structure in the region of the SD sequence; and fusion mRNAs consisting of the 5'-untranslated region of OppA mRNA and the open reading frame of beta-galactosidase. By measuring the synthesis of OppA or beta-galactosidase from these mRNAs, we found that the 171-nucleotide 5'-untranslated region and 145 nucleotides of the ORF of OppA mRNA are involved in the polyamine stimulation of OppA synthesis. When the secondary structure of the above region of OppA mRNA was analyzed by optimal computer folding, it was shown that the degree of polyamine stimulation of OppA protein synthesis was dependent on the structure of the SD sequence in addition to its position. Loose base pairing of the SD sequence with other regions of the mRNA caused strong polyamine stimulation, while intense base pairing of the SD sequence with other regions of the mRNA resulted in insignificant or weak polyamine stimulation.