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. |