title: NDR1-Dependent Regulation of Kindlin-3 Controls High-Affinity LFA-1 Binding and Immune Synapse Organization.
abstract: Antigen-specific adhesion between T cells and antigen-presenting cells (APC) during the formation of the immunological synapse (IS) is mediated by LFA-1 and ICAM-1. Here, LFA-1-ICAM-1 interactions were measured at the single-molecule level on supported lipid bilayers. High-affinity binding was detected at low frequencies in the inner peripheral supramolecular activation cluster (SMAC) zone that contained high levels of activated Rap1 and kindlin-3. Rap1 was essential for T cell attachment, whereas deficiencies of ste20-like kinases, Mst1/Mst2, diminished high-affinity binding and abrogated central SMAC (cSMAC) formation with mislocalized kindlin-3 and vesicle transport regulators involved in T cell receptor recycling/releasing machineries, resulting in impaired T cell-APC interactions. We found that NDR1 kinase, activated by the Rap1 signaling cascade through RAPL and Mst1/Mst2, associated with and recruited kindlin-3 to the IS, which was required for high-affinity LFA-1/ICAM-1 binding and cSMAC formation. Our findings reveal crucial roles for Rap1 signaling via NDR1 for recruitment of kindlin-3 and IS organization.
authors.affiliation: Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Naoyuki
authors.identifier: http://orcid.org/0000-0002-6624-5989
authors.initials: N
authors.lastname: Kondo
authors.affiliation: Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ueda
authors.affiliation: Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kita
authors.affiliation: Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Madoka
authors.initials: M
authors.lastname: Ozawa
authors.affiliation: Division of Gastroenterology and Hepatology, Third Department of Internal Medicine, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Tomiyama
authors.affiliation: Department of Urology and Andrology, Kansai Medical University, Hirakata, Osaka, Japan.
authors.forename: Kaneki
authors.initials: K
authors.lastname: Yasuda
authors.affiliation: Department of Biological Sciences, National Creative Research Initiatives Center, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, South Korea.
authors.forename: Dae-Sik
authors.initials: DS
authors.lastname: Lim
authors.affiliation: Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka, Japan kinashi@takii.kmu.ac.jp.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Kinashi
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D057809:HEK293 Cells; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D054992:Immunological Synapses / Q000378:metabolism*; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D000090742:Serine-Threonine Kinase 3; D015398:Signal Transduction; D000072760:Single Molecule Imaging; D022161:Transport Vesicles / Q000378:metabolism; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism; D000099259:N-myc Downstream-Regulated Gene 1 Protein; D000099317:Hippo Kinases
chemical_list: D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D017228:Hepatocyte Growth Factor; D018799:Intercellular Adhesion Molecule-1; D047908:Intracellular Signaling Peptides and Proteins; D016169:Lymphocyte Function-Associated Antigen-1; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D000090742:Serine-Threonine Kinase 3; D020827:rap1 GTP-Binding Proteins; D000099259:N-myc Downstream-Regulated Gene 1 Protein; C527818:kindlin-3 protein, mouse; C082332:macrophage stimulating protein; C490818:Stk3 protein, mouse; D000099317:Hippo Kinases
keywords: ICAM-1; LFA-1; NDR1; Rap1; immunological synapse; single-molecule measurement; vesicular transport