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To identify proteins acquiring a distinct phosphorylation status, herein, we compared the phosphorylation profile of macrophages upon exposure to Mtb and BCG. We observed a significant dephosphorylation of proteins following Mtb infection relative to those with uninfected or BCG-infected cells. A comprehensive tandem mass tag mass spectrometry (MS) approach detected \u223c10% phosphosites on a variety of host proteins that are modulated in response to infection. Interestingly, the innate immune-enhancing interferon (IFN)-stimulated genes were identified as a class of proteins differentially phosphorylated during infection, including the cytosolic RNA sensor RIG-I, which has been implicated in the immune response to bacterial infection. We show that Mtb infection results in the activation of RIG-I in primary human macrophages. Studies using RIG-I knockout macrophages reveal that the Mtb-mediated activation of RIG-I promotes IFN-\u03b2, IL-1\u03b1, and IL-1\u03b2 levels, dampens autophagy, and facilitates intracellular Mtb survival. To our knowledge, this is the first study providing exhaustive information on relative and quantitative changes in the global phosphoproteome profile of host macrophages that can be further explored in designing novel anti-TB drug targets. The peptide identification and MS\/MS spectra have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD013171.<\/jats:p>","DOI":"10.1021\/acs.jproteome.9b00895","type":"journal-article","created":{"date-parts":[[2020,5,14]],"date-time":"2020-05-14T18:25:52Z","timestamp":1589480752000},"page":"2316-2336","source":"Crossref","is-referenced-by-count":19,"title":["Relative and Quantitative\nPhosphoproteome Analysis\nof Macrophages in Response to Infection by Virulent and Avirulent\n                    <i>Mycobacteria<\/i>\n                    Reveals a Distinct Role of the Cytosolic RNA\nSensor RIG\u2011I in\n                    <i>Mycobacterium tuberculosis<\/i>\n                    Pathogenesis"],"prefix":"10.1021","volume":"19","author":[{"given":"Eira","family":"Choudhary","sequence":"first","affiliation":[{"name":"Translational Health Science and Technology Institute, NCR Biotech Science Cluster , , , ,","place":["Faridabad, Haryana, India, 121001"]},{"name":"Symbiosis International (Deemed University) , , , ,","place":["Pune, Maharashtra, India, 412115"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. Korin","family":"Bullen","sequence":"additional","affiliation":[{"name":"Johns Hopkins University , , , ,","place":["Baltimore, Maryland, United States, 21287"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renu","family":"Goel","sequence":"additional","affiliation":[{"name":"Translational Health Science and Technology Institute, NCR Biotech Science Cluster , , , ,","place":["Faridabad, Haryana, India, 121001"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alok Kumar","family":"Singh","sequence":"additional","affiliation":[{"name":"Johns Hopkins University , , , ,","place":["Baltimore, Maryland, United States, 21287"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monali","family":"Praharaj","sequence":"additional","affiliation":[{"name":"Johns Hopkins University , , , ,","place":["Baltimore, Maryland, United States, 21287"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Preeti","family":"Thakur","sequence":"additional","affiliation":[{"name":"Johns Hopkins University , , , ,","place":["Baltimore, Maryland, United States, 21287"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rohan","family":"Dhiman","sequence":"additional","affiliation":[{"name":"National Institute of Technology , , , ,","place":["Rourkela, Odisha, India, 769008"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William R.","family":"Bishai","sequence":"additional","affiliation":[{"name":"Johns Hopkins University , , , ,","place":["Baltimore, Maryland, United States, 21287"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9203-3026","authenticated-orcid":true,"given":"Nisheeth","family":"Agarwal","sequence":"additional","affiliation":[{"name":"Translational Health Science and Technology Institute, NCR Biotech Science Cluster , , , ,","place":["Faridabad, Haryana, India, 121001"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,5,14]]},"reference":[{"key":"2026081416162062000_cit1","doi-asserted-by":"publisher","first-page":"S2","DOI":"10.1038\/502S2a","article-title":"Epidemiology:\nA mortal foe","volume":"502","author":"Paulson","year":"2013","journal-title":"Nature"},{"key":"2026081416162062000_cit2","unstructured":"WHO\n          . 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