Title Influence of Acetobacter pasteurianus SKU1108 aspS Gene Expression on Escherichia coli Morphology
Author Kannipa Tasanapak1, Uraiwan Masud-Tippayasak1,2, Kazunobu Matsushita3, Wichien Yongmanitchai1, and Gunjana Theeragool1*
Address 1Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand, 2Center for Advanced Studies in Tropical Natural Resources, NRU-KU, Kasetsart University, Bangkok 10900, Thailand, 3Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi 753-8515, Japan
Bibliography Journal of Microbiology, 51(6),783–790, 2013,
DOI 10.1007/s12275-013-2619-6
Key Words Acetobacter pasteurianus, aspartyl-tRNA synthetase, aspS gene, ATP-binding domain, filamentous cell
Abstract The aspS gene encoding Aspartyl-tRNA synthetase (AspRS) from a thermotolerant acetic acid bacterium, Acetobacter pasteurianus SKU1108, has been cloned and characterized. The open reading frame (ORF) of the aspS gene consists of 1,788 bp, encoding 595 amino acid residues. The highly conserved Gly-Val-Asp-Arg ATP binding motif (motif 3) is located at the position 537-540 in the C-terminus. Deletion analysis of the aspS gene upstream region suggested that the promoter is around 173 bp upstream from the ATG initiation codon. Interestingly, transformation with the plasmids pGEM-T138, pUC138, and pCM138 synthesizing 138 amino acid C-terminal fragments of AspRS, that carry the ATP binding domain, caused E. coli cell lengthening at 37 and 42°C. Moreover, E. coli harboring pUC595 (synthesizing all 595 amino acids) and a disordered aspS gene in pGEM-T138 had normal rod shapes. The normal rod shape was observed in E. coli harboring pD539V following site-directed mutagenesis of the ATP binding domain. We propose that overproduction of truncated C-terminal peptides of AspRS may cause sequestration of intracellular ATP in E. coli, leaving less ATP for cell division or shaping cell morphology.