Structural basis of staphylococcal Stl inhibition on a eukaryotic dUTPase

Abstract

dUTPases are key enzymes in all life kingdoms. A staphylococcal repressor protein (Stl) inhibited dUTPases from multiple species to various extents. Understanding the molecular basis underlying the inhibition differences is crucial to develop effective proteinaceous inhibitors of dUTPases. Herein, we report the complex structure of Stl N-terminal domain (Stl^N-ter) and Litopenaeus vannamei dUTPase domain (lvDUT^65-210). Stl inhibited lvDUT^65-210 through its N-terminal domain. The lvDUT^65-210-Stl^N-ter complex structure revealed a heterohexamer encompassing three Stl^N-ter monomers bound to one lvDUT^65-210 trimer, generating two types of Stl-dUTPase interfaces. Interface I is formed by Stl interaction with the lvDUT^65-210 active-site region that is contributed by motifs I-IV from its two subunits; interface II results from Stl binding to the C-terminal motif V of the third lvDUT^65-210 subunit. Structural comparison revealed both conserved features and obvious differences in Stl-dUTPase interaction patterns, giving clues about the inhibition differences of Stl on dUTPases. Noticeably, interface II is only observed in lvDUT^65-210-Stl^N-ter. The Stl-interacting residues of lvDUT^65-210 are conserved in other eukaryotic dUTPases, particularly human dUTPase. Altogether, our study presents the first structural model of Stl interaction with eukaryotic dUTPase, contributing to a more complete view of Stl inhibition and facilitating the development of proteinaceous inhibitor for eukaryotic dUTPases.

Keywords: Crystal structure; Proteinaceous inhibitor; Stl dUTPase interactions.