ALBERT

Description: © The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Tarrant, E., P Riboldi, G., McIlvin, M. R., Stevenson, J., Barwinska-Sendra, A., Stewart, L. J., Saito, M. A., & Waldron, K. J. Copper stress in staphylococcus aureus leads to adaptive changes in central carbon metabolism. Metallomics, 11, (2019): 183-200, doi: 10.1039/C8MT00239H.

Description: Copper toxicity has been a long-term selection pressure on bacteria due to its presence in the environment and its use as an antimicrobial agent by grazing protozoa, by phagocytic cells of the immune system, and in man-made medical and commercial products. There is recent evidence that exposure to increased copper stress may have been a key driver in the evolution and spread of methicillin-resistant Staphylococcus aureus, a globally important pathogen that causes significant mortality and morbidity worldwide. Yet it is unclear how S. aureus physiology is affected by copper stress or how it adapts in order to be able to grow in the presence of excess copper. Here, we have determined quantitatively how S. aureus alters its proteome during growth under copper stress conditions, comparing this adaptive response in two different types of growth regime. We found that the adaptive response involves induction of the conserved copper detoxification system as well as induction of enzymes of central carbon metabolism, with only limited induction of proteins involved in the oxidative stress response. Further, we identified a protein that binds copper inside S. aureus cells when stressed by copper excess. This copper-binding enzyme, a glyceraldehyde-3-phosphate dehydrogenase essential for glycolysis, is inhibited by copper in vitro and inside S. aureus cells. Together, our data demonstrate that copper stress leads to the inhibition of glycolysis in S. aureus, and that the bacterium adapts to this stress by altering its central carbon utilisation pathways.

Description: KJW and ET were supported by a Sir Henry Dale Fellowship (to KJW) funded by the Wellcome Trust and the Royal Society (098375/Z/12/Z), and GPR was funded by a CAPES Science Without Borders scholarship (BEX 2445/13-1). JS (BB/F015895/1) and ABS (BB/J014516/1) were supported by BBSRC PhD studentships, and LS was supported by a Newcastle University Research Excellence Academy PhD studentship (05MREA). MAS was supported by the Gordon and Betty Moore Foundation Grant (GBM3782). We also thank Prof. Simon Foster (Sheffield, UK) for the gift of S. aureus strain SH1000, and Dr Julie Morrissey (Leicester, UK) for phage φ11.

Description: © The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Mazzotta, M. G., McIlvin, M. R., & Saito, M. A. Characterization of the Fe metalloproteome of a ubiquitous marine heterotroph, Pseudoalteromonas (BB2-AT2): multiple bacterioferritin copies enable significant Fe storage. Metallomics, (2020), doi:10.1039/d0mt00034e.

Description: Fe is a critical nutrient to the marine biological pump, which is the process that exports photosynthetically fixed carbon in the upper ocean to the deep ocean. Fe limitation controls photosynthetic activity in major regions of the oceans, and the subsequent degradation of exported photosynthetic material is facilitated particularly by marine heterotrophic bacteria. Despite their importance in the carbon cycle and the scarcity of Fe in seawater, the Fe requirements, storage and cytosolic utilization of these marine heterotrophs has been less studied. Here, we characterized the Fe metallome of Pseudoalteromonas (BB2-AT2). We found that with two copies of bacterioferritin (Bfr), Pseudoalteromonas possesses substantial capacity for luxury uptake of Fe. Fe : C in the whole cell metallome was estimated (assuming C : P stoichiometry ∼51 : 1) to be between ∼83 μmol : mol Fe : C, ∼11 fold higher than prior marine bacteria surveys. Under these replete conditions, other major cytosolic Fe-associated proteins were observed including superoxide dismutase (SodA; with other metal SOD isoforms absent under Fe replete conditions) and catalase (KatG) involved in reactive oxygen stress mitigation and aconitase (AcnB), succinate dehydrogenase (FrdB) and cytochromes (QcrA and Cyt1) involved in respiration. With the aid of singular value decomposition (SVD), we were able to computationally attribute peaks within the metallome to specific metalloprotein contributors. A putative Fe complex TonB transporter associated with the closely related Alteromonas bacterium was found to be abundant within the Pacific Ocean mesopelagic environment. Despite the extreme scarcity of Fe in seawater, the marine heterotroph Pseudoalteromonas has expansive Fe storage capacity and utilization strategies, implying that within detritus and sinking particles environments, there is significant opportunity for Fe acquisition. Together these results imply an evolved dedication of marine Pseudoalteromonas to maintaining an Fe metalloproteome, likely due to its dependence on Fe-based respiratory metabolism.

Description: M. G. M. was supported by the Camille and Henry Dreyfus Foundation Environmental Chemistry Postdoctoral Fellowship. We thank Kay Bidle (Rutgers University) for providing a culture of Pseudoalteromonas (BB2-AT2). We also thank Dawn Moran (WHOI) and Noelle Held (WHOI-MIT) for culturing assistance. We appreciate the Captain and Crew of the R/V Kilo Moana, and the many involved in the METZYME expedition sampling efforts. Discussions with Kevin Waldron (Newcastle University), Alison Butler (University of California, Santa Barbara), Lauren Manck (Scripps Institution of Oceanography), Randie Bundy (University of Washington) and Jake Gebbie (WHOI) were much appreciated. Funding for this research was provided by the Gordon and Betty Moore Foundation (3782), and NSF-OCE 1658030, 1736599, 1657766, 1924554, 1850719, 1924554.