A comprehensive two-hybrid analysis to explore the Legionella pneumophila effector-effector interactome.

Autor:	Mount HO; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada., Urbanus ML; Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada., Sheykhkarimli D; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada., Coté AG; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada., Laval F; Center for Cancer Systems Biology (CCSB), Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; TERRA Teaching and Research Centre, University of Liège, Gembloux, Belgium.; Laboratory of Viral Interactomes, GIGA Institute, University of Liège, Liège, Belgium.; Laboratory of Molecular and Cellular Epigenetics, GIGA Institute, University of Liège, Liège, Belgium., Coppin G; Center for Cancer Systems Biology (CCSB), Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; TERRA Teaching and Research Centre, University of Liège, Gembloux, Belgium., Kishore N; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada., Li R; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada., Spirohn-Fitzgerald K; Center for Cancer Systems Biology (CCSB), Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA., Petersen MO; Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.; Department of Microbiology and Immunology, The University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia., Knapp JJ; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada., Kim D-K; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada., Twizere J-C; TERRA Teaching and Research Centre, University of Liège, Gembloux, Belgium.; Laboratory of Viral Interactomes, GIGA Institute, University of Liège, Liège, Belgium., Calderwood MA; Center for Cancer Systems Biology (CCSB), Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA.; Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA., Vidal M; Center for Cancer Systems Biology (CCSB), Dana-Farber Cancer Institute, Boston, Massachusetts, USA.; Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, Massachusetts, USA., Roth FP; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.; Donnelly Centre, University of Toronto, Toronto, Ontario, Canada.; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, Ontario, Canada.; Department of Computational and Systems Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA., Ensminger AW; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.; Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada.
Jazyk:	angličtina
Zdroj:	MSystems [mSystems] 2024 Dec 17; Vol. 9 (12), pp. e0100424. Date of Electronic Publication: 2024 Nov 11.
DOI:	10.1128/msystems.01004-24
Abstrakt:	Legionella pneumophila uses over 300 translocated effector proteins to rewire host cells during infection and create a replicative niche for intracellular growth. To date, several studies have identified L. pneumophila effectors that indirectly and directly regulate the activity of other effectors, providing an additional layer of regulatory complexity. Among these are "metaeffectors," a special class of effectors that regulate the activity of other effectors once inside the host. A defining feature of metaeffectors is direct, physical interaction with a target effector. Metaeffector identification, to date, has depended on phenotypes in heterologous systems and experimental serendipity. Using a multiplexed, recombinant barcode-based yeast two-hybrid technology we screened for protein-protein interactions among all L. pneumophila effectors and 28 components of the Dot/Icm type IV secretion system (>167,000 protein combinations). Of the 52 protein interactions identified by this approach, 44 are novel protein interactions, including 10 novel effector-effector interactions (doubling the number of known effector-effector interactions). Importance: Secreted bacterial effector proteins are typically viewed as modulators of host activity, entering the host cytosol to physically interact with and modify the activity of one or more host proteins in support of infection. A growing body of evidence suggests that a subset of effectors primarily function to modify the activities of other effectors inside the host. These "effectors of effectors" or metaeffectors are often identified through experimental serendipity during the study of canonical effector function against the host. We previously performed the first global effector-wide genetic interaction screen for metaeffectors within the arsenal of Legionella pneumophila , an intracellular bacterial pathogen with over 300 effectors. Here, using a high-throughput, scalable methodology, we present the first global interaction network of physical interactions between L. pneumophila effectors. This data set serves as a complementary resource to identify and understand both the scope and nature of non-canonical effector activity within this important human pathogen. Competing Interests: F.P.R. and M.V. are advisors and shareholders of SeqWell, Inc. (Beverly, MA, USA).
Databáze:	MEDLINE
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