aureus has led to the search for alternative drug targets. Amongst them, proteins indispensable for cellular viability are optimal candidates. There are currently about 15 essential proteins from bacterial

genomes used as antibiotic targets encompassing a restricted set of microbial processes, including DNA replication and repair, fatty acid and protein biosynthesis, and cell wall synthesis [5]. A large number of essential proteins remain to be investigated for novel antimicrobial development. In a genome-wide study in Bacillus subtilis the IPTG-inducible Pspac conditional expression system was used to determine gene essentiality [6]. A subset of 15 genes identified in this screening had no significant homology to any gene of known function, and included the well-conserved Era/Obg family

of GTP PI3K inhibitor binding proteins [6]. The latter belongs to a diverse superfamily of the often referred to as low molecular weight GTPases, which act as molecular switches in the MCC950 solubility dmso regulation of crucial cellular processes across all domains of life, including: intracellular and membrane signalling, vesicular transport, cell division, chromosome partitioning, protein targeting and ribosomal function [7]. Although very few of the bacterial low molecular weight GTPases have well characterised roles, there is increasing evidence that members of the Era/Obg family of GTPases are involved in ribosome function, assembly or stability. Work on Era, Obg, YjeQ/YloQ, YlqF, YphC, and YsxC in E. coli and B. subtilis has indicated associations of these proteins Anlotinib nmr with ribosomal subunits and changes in ribosomal profiles [8–10]. Ribosome profiles, created by separation of ribosome constituents on a sucrose gradient, show a decrease in whole 70 S ribosomes with an concomitant increase in 30 S and 50 S ribosomal subunits after

depletion of the protein of interest [9, 11–15]. YsxC in B. subtilis (YihA in E. coli) is an ortholog of the Era/Obg family of GTP-binding protein CYTH4 that has been reported to be essential in B. subtilis, E. coli, S. pneumoniae, H. influenzae, and M. genitalium [9, 16, 17]. We have previously solved the crystal structure of the B. subtilis YsxC in its open and closed conformations, proven its ability to complex with GDP and GTP, and shown the conformational changes occurring upon nucleotide binding and GTP hydrolysis [18]. A B. subtilis mutant with ysxC under the control of the regulatable Pspank promoter has revealed that depletion of the protein led to the accumulation of intermediate 50 S subunits (described as 44.5 S subunits) different from those seen upon depletion of similar GTPases YphC and YlqF [9]. However, as with YlqF and YphC depletion, intermediates lacked ribosomal proteins L16, L36 and possibly L27. Other putative ribosomal interacting partners of YsxC have been suggested by Wicker-Planquart and co-authors [10]. YsxC is likely to be essential across eubacteria. In this study we demonstrate that YsxC of S.