title: Unbiased proteomic profiling strategy for discovery of bacterial effector proteins reveals that Salmonella protein PheA is a host cell cycle regulator.
abstract: Salmonella utilizes a type III secretion system to inject bacterial effector proteins into the host cell cytosol. Once in the cytosol, these effectors hijack various biochemical pathways to regulate virulence. Despite the importance of effector proteins, especially for understanding host-pathogen interactions, a potentially large number of effectors are yet to be identified. Here, we demonstrate that unbiased chemical proteomic profiling using off-the-shelf fluorescent probes leads to the discovery of a host cell cycle regulator encoded in the Salmonella genome. Our profiling combined with bioinformatic analysis implicates 29 Salmonella as potential effectors. We follow up on the top candidate, chorismate mutase-P/prehenate dehydratase, PheA, and present evidence that PheA is an effector that mimics E2F7 transcription factor of the host cell and promotes G1/S cell cycle arrest. This validates our strategy and opens opportunities for effector identification in the future.
authors: Molecular Recognition Research Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seoul 136-791, South Korea; Department of Biological Chemistry, University of Science & Technology, 113 Gwahank-ro, Yuseong-gu, Daejeon 305-333, South Korea.
authors: Ha-Na
authors: HN
authors: Na
authors: Molecular Recognition Research Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seoul 136-791, South Korea.
authors: Young-Hwa
authors: YH
authors: Yoo
authors: Molecular Recognition Research Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seoul 136-791, South Korea.
authors: Chang No
authors: CN
authors: Yoon
authors: Molecular Recognition Research Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seoul 136-791, South Korea; Department of Biological Chemistry, University of Science & Technology, 113 Gwahank-ro, Yuseong-gu, Daejeon 305-333, South Korea. Electronic address: junseoklee@kist.re.kr.
authors: Jun-Seok
authors: JS
authors: Lee
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D002460:Cell Line; D050702:E2F7 Transcription Factor / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D059585:G1 Phase Cell Cycle Checkpoints; D016680:Genome, Bacterial; D054884:Host-Pathogen Interactions; D051379:Mice; D008856:Microscopy, Fluorescence; D011302:Prephenate Dehydratase / Q000378:metabolism*; D020543:Proteome / Q000032:analysis; D040901:Proteomics; D059807:S Phase Cell Cycle Checkpoints; D012480:Salmonella Infections / Q000382:microbiology* / Q000503:physiopathology; D012486:Salmonella typhimurium / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D050702:E2F7 Transcription Factor; D005456:Fluorescent Dyes; D020543:Proteome; D011302:Prephenate Dehydratase