The Coxiella burnetii T4SS effector protein AnkG hijacks the 7SK small nuclear ribonucleoprotein complex for reprogramming host cell transcription

• Published in: PLoS pathogens Vol. 18; no. 2; p. e1010266
• Main Authors: Cordsmeier, Arne, Rinkel, Sven, Jeninga, Myriam, Schulze-Luehrmann, Jan, Ölke, Martha, Schmid, Benedikt, Hasler, Daniele, Meister, Gunter, Häcker, Georg, Petter, Michaela, Beare, Paul A, Lührmann, Anja
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.02.2022; Public Library of Science (PLoS)
• Subjects: Aerosols; Amino acids; Analysis; Antibodies; Apoptosis; Asymptomatic; Bacteria; Bacterial diseases; Bacterial proteins; Bacterial Proteins - metabolism; Biology and Life Sciences; Cell Survival; Cell viability; Coxiella burnetii; Coxiella burnetii - genetics; Coxiella burnetii - metabolism; Cytosol; DEAD-box RNA Helicases - metabolism; DNA helicase; DNA-directed RNA polymerase; Elongation; Endocarditis; Etiology; Experiments; Fever; Gene expression; Gene sequencing; Genetic transcription; Health aspects; HEK293 Cells; HeLa Cells; Host-Pathogen Interactions; Humans; Immunoprecipitation; Infections; Intracellular; Localization; Medicine and Health Sciences; Mutation; Pathogenesis; Pathogens; Physiological aspects; Positive Transcriptional Elongation Factor B - metabolism; Proteins; Q fever; Q Fever - metabolism; Q Fever - microbiology; Research and analysis methods; Ribonucleic acid; Ribonucleoproteins; Ribonucleoproteins (small nuclear); Ribonucleoproteins, Small Nuclear - metabolism; Rickettsia; RNA; RNA helicase; RNA polymerase; THP-1 Cells; Transcription elongation factor b; Type IV Secretion Systems - metabolism; Zoonoses; Germany
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1010266

Inhibition of host cell apoptosis is crucial for survival and replication of several intracellular bacterial pathogens. To interfere with apoptotic pathways, some pathogens use specialized secretion systems to inject bacterial effector proteins into the host cell cytosol. One of these pathogens is the obligate intracellular bacterium Coxiella burnetii, the etiological agent of the zoonotic disease Q fever. In this study, we analyzed the molecular activity of the anti-apoptotic T4SS effector protein AnkG (CBU0781) to understand how C. burnetii manipulates host cell viability. We demonstrate by co- and RNA-immunoprecipitation that AnkG binds to the host cell DExD box RNA helicase 21 (DDX21) as well as to the host cell 7SK small nuclear ribonucleoprotein (7SK snRNP) complex, an important regulator of the positive transcription elongation factor b (P-TEFb). The co-immunoprecipitation of AnkG with DDX21 is probably mediated by salt bridges and is independent of AnkG-7SK snRNP binding, and vice versa. It is known that DDX21 facilitates the release of P-TEFb from the 7SK snRNP complex. Consistent with the documented function of released P-TEFb in RNA Pol II pause release, RNA sequencing experiments confirmed AnkG-mediated transcriptional reprogramming and showed that expression of genes involved in apoptosis, trafficking, and transcription are influenced by AnkG. Importantly, DDX21 and P-TEFb are both essential for AnkG-mediated inhibition of host cell apoptosis, emphasizing the significance of the interaction of AnkG with both, the DDX21 protein and the 7SK RNA. In line with a critical function of AnkG in pathogenesis, the AnkG deletion C. burnetii strain was severely affected in its ability to inhibit host cell apoptosis and to generate a replicative C. burnetii-containing vacuole. In conclusion, the interference with the activity of regulatory host cell RNAs mediated by a bacterial effector protein represent a novel mechanism through which C. burnetii modulates host cell transcription, thereby enhancing permissiveness to bacterial infection.