Host glutathione is required for Rickettsia parkeri to properly septate, avoid ubiquitylation, and survive in macrophages.

Spotted fever group Rickettsia obligately reside in the cytosol where they parasitize over fifty metabolites from their hosts. However, the role for metabolite acquisition in pathogenesis remains unclear. Here, we find that depletion of the abundant low molecular weight thiol glutathione led to an impaired ability of Rickettsia parkeri to form plaques. Super-resolution microscopy revealed that glutathione depletion with buthionine sulfoximine (BSO) in endothelial or epithelial cells led to the formation of bacterial chains that increased in length over time. Chained bacteria had fewer actin-tails and were impeded for their ability to spread from cell to cell. Glutathione depletion also caused an increased frequency of colocalization between the bacterial surface and polyubiquitin. R. parkeri was significantly more restricted upon glutathione depletion in primary macrophages than in epithelial cells in a mechanism that avoided activating the inflammasome and the production of type I interferon. Together, these data suggest that host glutathione is critical for rickettsial septation, actin-based motility, avoiding ubiquitylation, and survival in immune cells.