The catalytic domain of pp56(lck), but not its regulatory domain, is sufficient for inducing IL-2 production.
The lymphoid src kinase pp56(lck) has been shown to be essential for the induction of different T lymphocyte responses, including CD4-mediated enhancement of Ag-induced T cell activation, early T cell differentiation, induction of IL-2 production, and cytotoxicity. It is assumed that pp56(lck) acts on these processes by phosphorylating substrates. However, it has been recently reported that the NH2 regulatory domain is sufficient to mediate CD4 accessory function. In this report we address the contribution of the regulatory and catalytic domains of pp56(lck) to another function of this enzyme independent of CD4: TCR-induced IL-2 production. Two pp56(lck) mutants lacking either the entire catalytic domain or the entire NH2 regulatory domain were generated, and their abilities to trigger transactivation of the TCR-regulated nuclear factor of activated T cells (NF-AT) region of the IL-2 promoter were compared. Only the catalytic, but not the NH2 regulatory, domain of pp56(lck) was able to induce NF-AT region transactivation on its own and to cooperate with other intracellular signals to trigger this response. Moreover, the catalytic domain of pp56(lck) was able to induce IL-2 cytokine production to an extent similar to that of wild-type pp56(lck). We conclude that different domains of the pp56(lck) molecule contribute to regulate distinct biologic functions. In fact, while the NH2 regulatory domain is sufficient to mediate CD4 accessory function, we show here that the catalytic domain of pp56(lck) is sufficient for induction of IL-2 production, mimicking TCR ligation.
National Center of Biotechnology, Autonoma University, Madrid, Spain.
A C
AC
Carrera
V
V
Calvo
L R
LR
Borlado
H
H
Paradis
S
S
Alemany
T M
TM
Roberts
C
C
Martinez-A
D000495:Allosteric Site; D001665:Binding Sites; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D004268:DNA-Binding Proteins / Q000502:physiology; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D015759:Ionomycin / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000473:pathology; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D050778:NFATC Transcription Factors; D009363:Neoplasm Proteins / Q000502:physiology; D009687:Nuclear Proteins; D010446:Peptide Fragments / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011948:Receptors, Antigen, T-Cell / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014157:Transcription Factors / Q000502:physiology; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D019061:src-Family Kinases / Q000737:chemistry / Q000502:physiology*
D004268:DNA-Binding Proteins; D007376:Interleukin-2; D007476:Ionophores; D050778:NFATC Transcription Factors; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D014157:Transcription Factors; D015759:Ionomycin; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D019061:src-Family Kinases; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium