title: Dispensability of zinc and the putative zinc-binding domain in bacterial glutamyl-tRNA synthetase.
abstract: The putative zinc-binding domain (pZBD) in Escherichia coli glutamyl-tRNA synthetase (GluRS) is known to correctly position the tRNA acceptor arm and modulate the amino acid-binding site. However, its functional role in other bacterial species is not clear since many bacterial GluRSs lack a zinc-binding motif in the pZBD. From experimental studies on pZBD-swapped E. coli GluRS, with Thermosynechoccus elongatus GluRS, Burkholderia thailandensis GluRS and E. coli glutamyl-queuosine-tRNA(Asp) synthetase (Glu-Q-RS), we show that E. coli GluRS, containing the zinc-free pZBD of B. thailandensis, is as functional as the zinc-bound wild-type E. coli GluRS, whereas the other constructs, all zinc-bound, show impaired function. A pZBD-tinkered version of E. coli GluRS that still retained Zn-binding capacity, also showed reduced activity. This suggests that zinc is not essential for the pZBD to be functional. From extensive structural and sequence analyses from whole genome database of bacterial GluRS, we further show that in addition to many bacterial GluRS lacking a zinc-binding motif, the pZBD is actually deleted in some bacteria, all containing either glutaminyl-tRNA synthetase (GlnRS) or a second copy of GluRS (GluRS2). Correlation between the absence of pZBD and the occurrence of glutamine amidotransferase CAB (GatCAB) in the genome suggests that the primordial role of the pZBD was to facilitate transamidation of misacylated Glu-tRNA(Gln) via interaction with GatCAB, whereas its role in tRNA(Glu) interaction may be a consequence of the presence of pZBD.
authors: *Department of Biophysics, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata 700054, India.
authors: Nipa
authors: N
authors: Chongdar
authors: *Department of Biophysics, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata 700054, India.
authors: Saumya
authors: S
authors: Dasgupta
authors: †Department of Biochemistry, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata 700054, India.
authors: Ajit Bikram
authors: AB
authors: Datta
authors: *Department of Biophysics, Bose Institute, P-1/12 CIT Scheme VIIM, Kolkata 700054, India.
authors: Gautam
authors: G
authors: Basu
mesh_terms: D019117:Burkholderia / Q000201:enzymology / Q000235:genetics; D030541:Databases, Genetic; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics; D016680:Genome, Bacterial; D005975:Glutamate-tRNA Ligase / Q000737:chemistry / Q000235:genetics; D017434:Protein Structure, Tertiary; D015032:Zinc / Q000737:chemistry*
chemical_list: D029968:Escherichia coli Proteins; D005975:Glutamate-tRNA Ligase; D015032:Zinc