title: Calmodulin and PI(3,4,5)P₃ cooperatively bind to the Itk pleckstrin homology domain to promote efficient calcium signaling and IL-17A production.
abstract: Precise regulation of the kinetics and magnitude of Ca(2+) signaling enables this signal to mediate diverse responses, such as cell migration, differentiation, vesicular trafficking, and cell death. We showed that the Ca(2+)-binding protein calmodulin (CaM) acted in a positive feedback loop to potentiate Ca(2+) signaling downstream of the Tec kinase family member Itk. Using NMR (nuclear magnetic resonance), we mapped CaM binding to two loops adjacent to the lipid-binding pocket within the Itk pleckstrin homology (PH) domain. The Itk PH domain bound synergistically to Ca(2+)/CaM and the lipid phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3], such that binding to Ca(2+)/CaM enhanced the binding to PI(3,4,5)P3 and vice versa. Disruption of CaM binding attenuated Itk recruitment to the membrane and diminished release of Ca(2+) from the endoplasmic reticulum. Moreover, disruption of this feedback loop abrogated Itk-dependent production of the proinflammatory cytokine IL-17A (interleukin-17A) by CD4(+) T cells. Additionally, we found that CaM associated with PH domains from other proteins, indicating that CaM may regulate other PH domain-containing proteins.
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, IA 50011, USA.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Boyken
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Jiancheng
authors.initials: J
authors.lastname: Hu
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Ryan P
authors.initials: RP
authors.lastname: Rimer
authors.affiliation: Department of Biochemistry, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Madeline A
authors.initials: MA
authors.lastname: Shea
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Andrey S
authors.initials: AS
authors.lastname: Shaw
authors.affiliation: Department of Biochemistry, Biophysics & Molecular Biology, Iowa State University, Ames, IA 50011, USA.
authors.forename: Amy H
authors.initials: AH
authors.lastname: Andreotti
authors.affiliation: Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA. Department of Pathology, Geisel School of Medicine at Dartmouth, Lebanon, NH 03756, USA. Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Lebanon, NH 03756, USA. yina.h.huang@dartmouth.edu.
authors.forename: Yina H
authors.initials: YH
authors.lastname: Huang
mesh_terms: D000818:Animals; D053898:Biosynthetic Pathways / Q000235:genetics / Q000502:physiology*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D020013:Calcium Signaling / Q000235:genetics / Q000502:physiology*; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015151:Immunoblotting; D020381:Interleukin-17 / Q000096:biosynthesis*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D010716:Phosphatidylinositols / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002147:Calmodulin; D020381:Interleukin-17; D010716:Phosphatidylinositols; C118303:phosphoinositide-3,4,5-triphosphate; D011505:Protein-Tyrosine Kinases; C080813:emt protein-tyrosine kinase
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