Letter | Published:

Prophage WO genes recapitulate and enhance Wolbachia-induced cytoplasmic incompatibility

Nature volume 543, pages 243247 (09 March 2017) | Download Citation


The genus Wolbachia is an archetype of maternally inherited intracellular bacteria that infect the germline of numerous invertebrate species worldwide. They can selfishly alter arthropod sex ratios and reproductive strategies to increase the proportion of the infected matriline in the population. The most common reproductive manipulation is cytoplasmic incompatibility, which results in embryonic lethality in crosses between infected males and uninfected females. Females infected with the same Wolbachia strain rescue this lethality. Despite more than 40 years of research1 and relevance to symbiont-induced speciation2,3, as well as control of arbovirus vectors4,5,6 and agricultural pests7, the bacterial genes underlying cytoplasmic incompatibility remain unknown. Here we use comparative and transgenic approaches to demonstrate that two differentially transcribed, co-diverging genes in the eukaryotic association module of prophage WO8 from Wolbachia strain wMel recapitulate and enhance cytoplasmic incompatibility. Dual expression in transgenic, uninfected males of Drosophila melanogaster crossed to uninfected females causes embryonic lethality. Each gene additively augments embryonic lethality in crosses between infected males and uninfected females. Lethality associates with embryonic defects that parallel those of wild-type cytoplasmic incompatibility and is notably rescued by wMel-infected embryos in all cases. The discovery of cytoplasmic incompatibility factor genes cifA and cifB pioneers genetic studies of prophage WO-induced reproductive manipulations and informs the continuing use of Wolbachia to control dengue and Zika virus transmission to humans.

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This work was supported by National Institutes of Health (NIH) R21 HD086833 and National Science Foundation IOS 1456778 to Seth R.B., National Science Foundation DEB-1501398 and NIH 5T32GM008554 training grant support to D.P.L., NIH T32GM07347 training grant support for J.A.M. to the Vanderbilt Medical Scientist Training Program, and NIH AI081322 to A.M.F. Imaging was performed in part through the use of the Vanderbilt University Medical Center Cell Imaging Shared Resource (supported by NIH grants CA68485, DK20593, DK58404, DK59637, and EY08126). We thank K. Jernigan and P. Snider for help with preliminary studies, and A. Brooks for assistance with figure preparation.

Author information

Author notes

    • Daniel P. LePage
    •  & Jason A. Metcalf

    These authors contributed equally to this work.


  1. Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee 37235, USA

    • Daniel P. LePage
    • , Jason A. Metcalf
    • , Sarah R. Bordenstein
    • , Jungmin On
    • , Jessamyn I. Perlmutter
    • , J. Dylan Shropshire
    • , Emily M. Layton
    • , Lisa J. Funkhouser-Jones
    •  & Seth R. Bordenstein
  2. Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520, USA

    • John F. Beckmann
  3. Department of Pathology, Microbiology, and Immunology, Vanderbilt University, Nashville, Tennessee 37235, USA

    • Seth R. Bordenstein


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D.P.L. performed gene expression and hatch rate assays, embryo cytology, and assayed for transgene and infection status of flies. J.A.M. performed comparative genomics analyses, generated transgenic flies, and drafted the manuscript. Sarah R.B. performed evolutionary and bioinformatic analyses. J.O. performed hatch rates, assayed sex ratios, collected flies for all experiments, and assayed for transgene and infection status of flies. J.I.P. conducted younger brother effect experiments and performed embryo cytology. J.D.S. performed hatch rate assays, collected flies for parallel embryo cytology, and assayed for transgene and infection status of flies. E.M.L. collected flies and performed hatch rate assays. L.J.F.-J. obtained the wVitA transcriptome. J.F.B. obtained the wPip proteome. Seth R.B. supervised the work and contributed to all experimental designs, data analysis, and data interpretation. All authors participated in manuscript preparation, editing, and final approval.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Seth R. Bordenstein.

Reviewer Information Nature thanks S. L. O’Neill, W. Sullivan and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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