Legionella effector AnkX interacts with host nuclear protein PLEKHN1.

Autor: Yu X; State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences-Beijing (PHOENIX Center), Beijing Institute of Radiation Medicine, Beijing, 102206, China., Noll RR; Department of Biological Sciences, University of Delaware, 105 The Green, Newark, DE, 19716, USA., Romero Dueñas BP; Department of Biological Sciences, University of Delaware, 105 The Green, Newark, DE, 19716, USA., Allgood SC; Department of Biological Sciences, University of Delaware, 105 The Green, Newark, DE, 19716, USA., Barker K; Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ, 85287, USA., Caplan JL; Department of Biological Sciences, University of Delaware, 105 The Green, Newark, DE, 19716, USA.; Delaware Biotechnology Institute, University of Delaware, Newark, 19716, DE, USA., Machner MP; Cell Biology and Neurobiology Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA., LaBaer J; Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ, 85287, USA., Qiu J; Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ, 85287, USA. Ji.Qiu@asu.edu., Neunuebel MR; Department of Biological Sciences, University of Delaware, 105 The Green, Newark, DE, 19716, USA. neunr@udel.edu.
Jazyk: angličtina
Zdroj: BMC microbiology [BMC Microbiol] 2018 Jan 05; Vol. 18 (1), pp. 5. Date of Electronic Publication: 2018 Jan 05.
DOI: 10.1186/s12866-017-1147-7
Abstrakt: Background: The intracellular bacterial pathogen Legionella pneumophila proliferates in human alveolar macrophages, resulting in a severe pneumonia termed Legionnaires' disease. Throughout the course of infection, L. pneumophila remains enclosed in a specialized membrane compartment that evades fusion with lysosomes. The pathogen delivers over 300 effector proteins into the host cell, altering host pathways in a manner that sets the stage for efficient pathogen replication. The L. pneumophila effector protein AnkX targets host Rab GTPases and functions in preventing fusion of the Legionella-containing vacuole with lysosomes. However, the current understanding of AnkX's interaction with host proteins and the means through which it exerts its cellular function is limited.
Results: Here, we investigated the protein interaction network of AnkX by using the nucleic acid programmable protein array (NAPPA), a high-density platform comprising 10,000 unique human ORFs. This approach facilitated the discovery of PLEKHN1 as a novel interaction partner of AnkX. We confirmed this interaction through multiple independent in vitro pull-down, co-immunoprecipitation, and cell-based assays. Structured illumination microscopy revealed that endogenous PLEKHN1 is found in the nucleus and on vesicular compartments, whereas ectopically produced AnkX co-localized with lipid rafts at the plasma membrane. In mammalian cells, HaloTag-AnkX co-localized with endogenous PLEKHN1 on vesicular compartments. A central fragment of AnkX (amino acids 491-809), containing eight ankyrin repeats, extensively co-localized with endogenous PLEKHN1, indicating that this region may harbor a new function. Further, we found that PLEKHN1 associated with multiple proteins involved in the inflammatory response.
Conclusions: Altogether, our study provides evidence that in addition to Rab GTPases, the L. pneumophila effector AnkX targets nuclear host proteins and suggests that AnkX may have novel functions related to manipulating the inflammatory response.
Databáze: MEDLINE
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