title: The Tec Kinase-Regulated Phosphoproteome Reveals a Mechanism for the Regulation of Inhibitory Signals in Murine Macrophages.
abstract: Previous work has shown conflicting roles for Tec family kinases in regulation of TLR-dependent signaling in myeloid cells. In the present study, we performed a detailed investigation of the role of the Tec kinases Btk and Tec kinases in regulating TLR signaling in several types of primary murine macrophages. We demonstrate that primary resident peritoneal macrophages deficient for Btk and Tec secrete less proinflammatory cytokines in response to TLR stimulation than do wild-type cells. In contrast, we found that bone marrow-derived and thioglycollate-elicited peritoneal macrophages deficient for Btk and Tec secrete more proinflammatory cytokines than do wild-type cells. We then compared the phosphoproteome regulated by Tec kinases and LPS in primary peritoneal and bone marrow-derived macrophages. From this analysis we determined that Tec kinases regulate different signaling programs in these cell types. In additional studies using bone marrow-derived macrophages, we found that Tec and Btk promote phosphorylation events necessary for immunoreceptor-mediated inhibition of TLR signaling. Taken together, our results are consistent with a model where Tec kinases (Btk, Tec, Bmx) are required for TLR-dependent signaling in many types of myeloid cells. However, our data also support a cell type-specific TLR inhibitory role for Btk and Tec that is mediated by immunoreceptor activation and signaling via PI3K.
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101;
authors.forename: Giacomo
authors.initials: G
authors.lastname: Tampella
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101;
authors.forename: Hannah M
authors.initials: HM
authors.lastname: Kerns
authors.affiliation: Celgene Avilomics Research, Bedford, MA 01730;
authors.forename: Deqiang
authors.initials: D
authors.lastname: Niu
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101;
authors.forename: Swati
authors.initials: S
authors.lastname: Singh
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101;
authors.forename: Socheath
authors.initials: S
authors.lastname: Khim
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101;
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Bosch
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101;
authors.forename: Meghan E
authors.initials: ME
authors.lastname: Garrett
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101; Department of Immunology, University of Washington School of Medicine, Seattle WA 98195;
authors.forename: Albanus
authors.initials: A
authors.lastname: Moguche
authors.affiliation: Celgene Avilomics Research, Bedford, MA 01730;
authors.forename: Erica
authors.initials: E
authors.lastname: Evans
authors.affiliation: Celgene Avilomics Research, Bedford, MA 01730;
authors.forename: Beth
authors.initials: B
authors.lastname: Browning
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle WA 98195;
authors.forename: Tahmina A
authors.initials: TA
authors.lastname: Jahan
authors.affiliation: Celgene Avilomics Research, Bedford, MA 01730;
authors.forename: Mariana
authors.initials: M
authors.lastname: Nacht
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle WA 98195;
authors.forename: Alejandro
authors.initials: A
authors.lastname: Wolf-Yadlin
authors.affiliation: Experimental Sciences, Pediatrics Clinic and Institute for Molecular Medicine A. Nocivelli, University of Brescia, Civil Hospital of Brescia, 25100 Bescia, Italy;
authors.forename: Alessandro
authors.initials: A
authors.lastname: Plebani
authors.affiliation: Department of Immunology, University of Washington School of Medicine, Seattle WA 98195; Benaroya Research Institute, Seattle WA 98101; and.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Hamerman
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101; Department of Immunology, University of Washington School of Medicine, Seattle WA 98195; Department of Pediatrics, University of Washington School of Medicine, Seattle WA 98195 drawling@uw.edu rickerj@uw.edu.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rawlings
authors.affiliation: Seattle Children's Research Institute, Seattle WA 98101; Department of Pediatrics, University of Washington School of Medicine, Seattle WA 98195 drawling@uw.edu rickerj@uw.edu.
authors.forename: Richard G
authors.initials: RG
authors.lastname: James
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009928:Organ Specificity; D010529:Peritoneal Cavity / Q000166:cytology; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000276:immunology; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology*; D010766:Phosphorylation; D061251:Primary Cell Culture; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000276:immunology*; D015398:Signal Transduction; D051193:Toll-Like Receptors / Q000235:genetics / Q000276:immunology
chemical_list: D008070:Lipopolysaccharides; D010750:Phosphoproteins; D051193:Toll-Like Receptors; C492366:Bmx protein, mouse; D019869:Phosphatidylinositol 3-Kinases; C089848:Tec protein-tyrosine kinase; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase