Structural basis for (p)ppGpp synthesis by the Staphylococcus aureus small alarmone synthetase RelP.
Melek Cemre ManavJelena BeljantsevaMartin S BojerTanel TensonHanne IngmerVasili HauryliukDitlev E BrodersenPublished in: The Journal of biological chemistry (2018)
The stringent response is a global reprogramming of bacterial physiology that renders cells more tolerant to antibiotics and induces virulence gene expression in pathogens in response to stress. This process is driven by accumulation of the intracellular alarmone guanosine-5'-di(tri)phosphate-3'-diphosphate ((p)ppGpp), which is produced by enzymes of the RelA SpoT homologue (RSH) family. The Gram-positive Firmicute pathogen, Staphylococcus aureus, encodes three RSH enzymes: a multidomain RSH (Rel) that senses amino acid starvation on the ribosome and two small alarmone synthetase (SAS) enzymes, RelQ (SAS1) and RelP (SAS2). In Bacillus subtilis, RelQ (SAS1) was shown to form a tetramer that is activated by pppGpp and inhibited by single-stranded RNA, but the structural and functional regulation of RelP (SAS2) is unexplored. Here, we present crystal structures of S. aureus RelP in two major functional states, pre-catalytic (bound to GTP and the non-hydrolyzable ATP analogue, adenosine 5'-(α,β-methylene)triphosphate (AMP-CPP)), and post-catalytic (bound to pppGpp). We observed that RelP also forms a tetramer, but unlike RelQ (SAS1), it is strongly inhibited by both pppGpp and ppGpp and is insensitive to inhibition by RNA. We also identified putative metal ion-binding sites at the subunit interfaces that were consistent with the observed activation of the enzyme by Zn2+ ions. The structures reported here reveal the details of the catalytic mechanism of SAS enzymes and provide a molecular basis for understanding differential regulation of SAS enzymes in Firmicute bacteria.
Keyphrases
- staphylococcus aureus
- gene expression
- biofilm formation
- structural basis
- amino acid
- bacillus subtilis
- gram negative
- escherichia coli
- protein kinase
- high resolution
- antimicrobial resistance
- induced apoptosis
- crystal structure
- genome wide
- nucleic acid
- oxidative stress
- heavy metals
- reactive oxygen species
- cell cycle arrest
- cystic fibrosis
- pi k akt
- stress induced