Coxiella burnetii encodes an LvgA‐related protein important for intracellular replication

Coxiella burnetii is a bacterial pathogen that replicates in a specialised lysosome‐derived organelle called the Coxiella‐containing vacuole (CCV). Establishment of the CCV requires the Dot/Icm type IVB secretion system. A previous transposon mutagenesis screen identified the gene cbu1754 as being important for the intracellular replication of C. burnetii. To understand the function of the protein encoded by cbu1754, CCV maturation and intracellular replication phenotypes of a cbu1754 mutant were analysed. In contrast to vacuoles containing wild‐type C. burnetii Nine Mile phase II, vacuoles containing the isogenic cbu1754 mutant were smaller and did not display detectible amounts of the autophagy protein LC3, which indicated a CCV biogenesis defect. The Cbu1754 protein was not efficiently delivered into the host cell cytosol during infection, which indicated this protein is not a Dot/Icm‐translocated effector protein. Secondary structure predictions suggested that Cbu1754 could be similar to the Legionella pneumophila LvgA protein, which is a component of the Dot/Icm apparatus. Consistent with this hypothesis, production of Cbu1754 in an L. pneumophila ∆lvgA mutant restored LvgA‐dependent activities. The L. pneumophila proteins LvgA, IcmS and IcmW are interacting partners that comprise a subassembly of the coupling protein complex that mediates Dot/Icm‐dependent effector translocation. Similarly, the Cbu1754 protein was found to be a component of the chaperone complex containing the C. burnetii proteins IcmS and IcmW. Thus, the Cbu1754 protein is an LvgA‐related protein important for Dot/Icm function and intracellular replication of C. burnetii. The intracellular bacterial pathogen Coxiella burnetii uses a Dot/Icm type IVB secretion system to deliver effector proteins into host cells during intracellular growth. The Cbu1754 protein was found to be important for the biogenesis of the Coxiella‐containing vacuole (CCV) and intracellular replication. Our data demonstrate that the Cbu1754 protein through interactions with IcmSW is a component of the Dot/Icm coupling protein complex, which is a type IVB subassembly essential for recruitment and delivery of effectors to the secretion machinery.