Vico-Barranco, InmaculadaArbulo-Echevarria, Mikel M.Serrano-García, IsabelPérez-Linaza, AlbaMiranda-Sayago, José M.Miazek, ArkadiuszNarbona-Sánchez, IsaacAguado, Enrique2022-08-292022-08-292021-02-06Vico-Barranco I, Arbulo-Echevarria MM, Serrano-García I, Pérez-Linaza A, Miranda-Sayago JM, Miazek A. A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling. Cells. 2021 Feb 6;10(2):343http://hdl.handle.net/10668/3963Intracellular signaling through the T cell receptor (TCR) is essential for T cell development and function. Proper TCR signaling requires the sequential activities of Lck and ZAP-70 kinases, which result in the phosphorylation of tyrosine residues located in the CD3 ITAMs and the LAT adaptor, respectively. LAT, linker for the activation of T cells, is a transmembrane adaptor protein that acts as a scaffold coupling the early signals coming from the TCR with downstream signaling pathways leading to cellular responses. The leukemic T cell line Jurkat and its derivative mutants J.CaM1.6 (Lck deficient) and J.CaM2 (LAT deficient) have been widely used to study the first signaling events upon TCR triggering. In this work, we describe the loss of LAT adaptor expression found in a subline of J.CaM1.6 cells and analyze cis-elements responsible for the LAT expression defect. This new cell subline, which we have called J.CaM1.7, can re-express LAT adaptor after Protein Kinase C (PKC) activation, which suggests that activation-induced LAT expression is not affected in this new cell subline. Contrary to J.CaM1.6 cells, re-expression of Lck in J.CaM1.7 cells was not sufficient to recover TCR-associated signals, and both LAT and Lck had to be introduced to recover activatory intracellular signals triggered after CD3 crosslinking. Overall, our work shows that the new LAT negative J.CaM1.7 cell subline could represent a new model to study the functions of the tyrosine kinase Lck and the LAT adaptor in TCR signaling, and their mutual interaction, which seems to constitute an essential early signaling event associated with the TCR/CD3 complex.enAtribución 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/LckLATJ.CaM1.6TCRSignalingT cell receptorPhosphorylationT lymphocytesProtein kinase CReceptores de antígenos de linfocitos TFosforilaciónLinfocitos TProteína quinasa CMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Intracellular Signaling Peptides and Proteins::Adaptor Proteins, Signal TransducingMedical Subject Headings::Information Science::Information Science::Information Services::Documentation::Molecular Sequence Data::Base SequenceMedical Subject Headings::Chemicals and Drugs::Biological Factors::Blood Coagulation Factors::CalciumMedical Subject Headings::Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell SurvivalMedical Subject Headings::Phenomena and Processes::Metabolic Phenomena::Metabolism::Enzyme ActivationMedical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::HumansMedical Subject Headings::Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, Tumor::Jurkat CellsMedical Subject Headings::Organisms::Viruses::RNA Viruses::Retroviridae::LentivirusMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Tyrosine Kinases::src-Family Kinases::Lymphocyte Specific Protein Tyrosine Kinase p56(lck)Medical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Membrane ProteinsMedical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Structures::PlasmidsMedical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Structures::Genome::Genome Components::Genes::Gene Components::Regulatory Elements, Transcriptional::Promoter Regions, GeneticMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Serine-Threonine Kinases::Protein Kinase CMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Membrane Proteins::Receptors, Cell Surface::Receptors, Immunologic::Receptors, Antigen::Receptors, Antigen, T-CellMedical Subject Headings::Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Signal TransductionMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Intracellular Signaling Peptides and Proteins::ZAP-70 Protein-Tyrosine KinaseMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Membrane Proteins::Receptors, Cell Surface::Receptors, Immunologic::Receptors, Antigen::Receptors, Antigen, T-Cell::Receptor-CD3 Complex, Antigen, T-CellMedical Subject Headings::Phenomena and Processes::Metabolic Phenomena::Metabolism::PhosphorylationMedical Subject Headings::Anatomy::Cells::Blood Cells::Leukocytes::Leukocytes, Mononuclear::Lymphocytes::T-LymphocytesMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Amino Acids::Amino Acids, Cyclic::Amino Acids, Aromatic::TyrosineMedical Subject Headings::Anatomy::Cells::Cells, Cultured::Cell Lineresearch article33562083open access10.3390/cells100203432073-4409PMC7915312