A Secondary Structure in the 5' Untranslated Region of adhE mRNA Required for RNase G-Dependent Regulation

• Published in: Bioscience, biotechnology, and biochemistry Vol. 77; no. 12; pp. 2473 - 2479
• Main Authors: ITO, Kazutaka, HAMASAKI, Kohshin, KAYAMORI, Aya, NGUYEN, Phuong Anh Thi, AMAGAI, Kaoru, WACHI, Masaaki
• Format: Journal Article
• Language: English
• Published: England Japan Society for Bioscience, Biotechnology, and Agrochemistry 2013; Oxford University Press
• Subjects: 5' Untranslated Regions - genetics; 5'-UTR; adhE; Alcohol Dehydrogenase - genetics; Aldehyde Oxidoreductases - genetics; Base Sequence; Binding Sites; Endoribonucleases - metabolism; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Gene Expression Regulation, Bacterial; Lac Operon - genetics; mRNA stability; Nucleic Acid Conformation; Protein Binding; RNase G
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.130618

Escherichia coli RNase G is involved in the degradation of several mRNAs, including adhE and eno, which encode alcohol dehydrogenase and enolase respectively. Previous research indicates that the 5' untranslated region (5'-UTR) of adhE mRNA gives RNase G-dependency to lacZ mRNA when tagged at the 5'-end, but it has not been elucidated yet how RNase G recognizes adhE mRNA. Primer extension analysis revealed that RNase G cleaved a phosphodiester bond between −19A and −18C in the 5'-UTR (the A of the start codon was defined as +1). Site-directed mutagenesis indicated that RNase G did not recognize the nucleotides at −19 and −18. Random deletion analysis indicated that the sequence from −145 to −125 was required for RNase G-dependent degradation. Secondary structure prediction and further site-directed deletion suggested that the stem-loop structure, with a bubble in the stem, is required for RNaseG-dependent degradation of adhE mRNA.