Crystal structure of a guanine nucleotide exchange factor encoded by the scrub typhus pathogen Orientia tsutsugamushi

Lim, Christopher; Berk, Jason M.; Blaise, Alyssa; Bircher, Josie; Koleske, Anthony J.; Hochstrasser, Mark; Xiong, Yong

doi:10.1073/pnas.2018163117

Title: Crystal structure of a guanine nucleotide exchange factor encoded by the scrub typhus pathogen Orientia tsutsugamushi

Journal Article · Thu Nov 12 00:00:00 EST 2020 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2018163117· OSTI ID:1734934

^[1]; Berk, Jason M. ^[1]; Blaise, Alyssa ^[1]; Bircher, Josie ^[1]; Koleske, Anthony J. ^[1]; Hochstrasser, Mark ^[1];

^[1]

Yale Univ., New Haven, CT (United States)

Rho family GTPases regulate an array of cellular processes and are often modulated by pathogens to promote infection. In this work, we identify a cryptic guanine nucleotide exchange factor (GEF) domain in the OtDUB protein encoded by the pathogenic bacterium Orientia tsutsugamushi. A proteomics-based OtDUB interaction screen identified numerous potential host interactors, including the Rho GTPases Rac1 and Cdc42. We discovered a domain in OtDUB with Rac1/Cdc42 GEF activity (OtDUB_GEF), with higher activity toward Rac1 in vitro. While this GEF bears no obvious sequence similarity to known GEFs, crystal structures of OtDUB_GEF alone (3.0 Å) and complexed with Rac1 (1.7 Å) reveal striking convergent evolution, with a unique topology, on a V-shaped bacterial GEF fold shared with other bacterial GEF domains. Structure-guided mutational analyses identified residues critical for activity and a mechanism for nucleotide displacement. Ectopic expression of OtDUB activates Rac1 preferentially in cells, and expression of the OtDUBGEF alone alters cell morphology. Cumulatively, this work reveals a bacterial GEF within the multifunctional OtDUB that co-opts host Rac1 signaling to induce changes in cytoskeletal structure.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-06CH11357; R01 AI116313; R01 GM046904; R35 GM136325; R01 MH115939; NS105640; R21 NS112121; DGE1122492; R01 MH115939-03S1; F31 NS113511

OSTI ID:: 1734934

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 117, Issue 48; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Orientia tsutsugamushi
x-ray crystallography
guanine nucleotide exchange
factor
scrub typhus
Rac1

Title: Crystal structure of a guanine nucleotide exchange factor encoded by the scrub typhus pathogen Orientia tsutsugamushi

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