Crystal structure of the YoeBSa1-YefMSa1 complex from Staphylococcus aureus

Abstract

Toxin-antitoxin (TA) systems are ubiquitously found in bacteria and are related to cell maintenance and survival under environmental stresses such as heat shock, nutrient starvation, and antibiotic treatment. Here, we report for the first time the crystal structure of the Staphylococcus aureus TA complex YoeB_Sa1-YefM_Sa1 at a resolution of 1.7 Å. This structure reveals a heterotetramer with a 2:2 stoichiometry between YoeB_Sa1 and YefM_Sa1. The N-terminal regions of the YefM_Sa1 antitoxin form a homodimer characteristic of a hydrophobic core, and the C-terminal extended region of each YefM_Sa1 protomer makes contact with each YoeB_Sa1 monomer. The binding stoichiometry of YoeB_Sa1 and YefM_Sa1 is different from that of YoeB and YefM of E. coli (YoeB_Ec and YefM_Ec), which is the only structural homologue among YoeB-YefM families; however, the structures of individual YoeB_Sa1 and YefM_Sa1 subunits in the complex are highly similar to the corresponding structures in E. coli. In addition, docking simulation with a minimal RNA substrate provides structural insight into the guanosine specificity of YoeB_Sa1 for cleavage in the active site, which is distinct from the specificity of YoeB_Ec for adenosine rather than guanosine. Given the previous finding that YoeB_Sa1 exhibits fatal toxicity without inducing persister cells, the structure of the YoeB_Sa1-YefM_Sa1 complex will contribute to the design of a new category of anti-staphylococcal agents that disrupt the YoeB_Sa1-YefM_Sa1 complex and increase YoeB_Sa1 toxicity.

Keywords: Staphylococcus aureus; Toxin – Antitoxin system; X-ray crystallography.