Wolbachia are obligate intracellular bacteria1 that infect arthropods, including approximately two-thirds of insect species2. Wolbachia manipulate insect reproduction by enhancing their inheritance through the female germline. The most common alteration is cytoplasmic incompatibility (CI)3–5, where eggs from uninfected females fail to develop when fertilized by sperm from Wolbachia-infected males. By contrast, if female and male partners are both infected, embryos are viable. CI is a gene-drive mechanism impacting population structure6 and causing reproductive isolation7, but its molecular mechanism has remained unknown. We show that a Wolbachia deubiquitylating enzyme (DUB) induces CI. The CI-inducing DUB, CidB, cleaves ubiquitin from substrates and is encoded in a two-gene operon, and the other protein, CidA, binds CidB. Binding is strongest between cognate partners in cidA-cidB homologues. In transgenic Drosophila, the cidA-cidB operon mimics CI when sperm introduce it into eggs, and a catalytically inactive DUB does not induce sterility. Toxicity is recapitulated in yeast by CidB alone; this requires DUB activity but is rescued by coexpressed CidA. A paralogous operon involves a putative nuclease (CinB) rather than a DUB. Analogous binding, toxicity and rescue in yeast were observed. These results identify a CI mechanism involving interacting proteins that are secreted into germline cells by Wolbachia, and suggest new methods for insect control.
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The authors thank A. Fallon, R. Tomko Jr and A. Chavez for critical review of the manuscript, L. Klasson for sharing preliminary data, H. Frydman and L. Cooley for fly lines, J. Carlson, K. Menuz, P. Graham and C. Yam for support with fly experimentation, C. Schlieker for use of instruments and C. Das and M. Sheedlo for HA-Ub-VME. Funding was provided by USDA-NIFA postdoctoral fellowship 2014-67012-22268 (to J.F.B.), the Marion Brooks-Wallace fellowship (to J.F.B.) and National Institutes of Health grant GM053756 (to M.H.).
The authors declare no competing financial interests.
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Beckmann, J., Ronau, J. & Hochstrasser, M. A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility. Nat Microbiol 2, 17007 (2017). https://doi.org/10.1038/nmicrobiol.2017.7
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