PMID- 10318843
OWN - NLM
STAT- MEDLINE
DCOM- 19990617
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 20
DP  - 1999 May 14
TI  - Tyrosine 319 in the interdomain B of ZAP-70 is a binding site for the Src
      homology 2 domain of Lck.
PG  - 14229-37
AB  - T-cell antigen receptor-induced signaling requires both ZAP-70 and Lck
      protein-tyrosine kinases. One essential function of Lck in this process is to
      phosphorylate ZAP-70 and up-regulate its catalytic activity. We have previously
      shown that after T-cell antigen receptor stimulation, Lck binds to ZAP-70 via its
      Src homology 2 (SH2) domain (LckSH2) and, more recently, that Tyr319 of ZAP-70 is
      phosphorylated in vivo and plays a positive regulatory role. Here, we
      investigated the possibility that Tyr319 mediates the SH2-dependent interaction
      between Lck and ZAP-70. We show that a phosphopeptide encompassing the motif
      harboring Tyr319, YSDP, interacted with LckSH2, although with a lower affinity
      compared with a phosphopeptide containing the optimal binding motif, YEEI.
      Moreover, mutation of Tyr319 to phenylalanine prevented the interaction of ZAP-70
      with LckSH2. Based on these results, a gain-of-function mutant of ZAP-70 was
      generated by changing the sequence Y319SDP into Y319EEI. As a result of its
      increased ability to bind LckSH2, this mutant induced a dramatic increase in NFAT
      activity in Jurkat T-cells, was hyperphosphorylated, and displayed a higher
      catalytic activity compared with wild-type ZAP-70. Collectively, our findings
      indicate that Tyr319-mediated binding of the SH2 domain of Lck is crucial for
      ZAP-70 activation and consequently for the propagation of the signaling cascade
      leading to T-cell activation.
FAU - Pelosi, M
AU  - Pelosi M
AD  - Molecular Immunology Unit, Institut Pasteur, 25-28 Rue du Docteur Roux, 75724
      Paris Cedex 15, France.
FAU - Di Bartolo, V
AU  - Di Bartolo V
FAU - Mounier, V
AU  - Mounier V
FAU - Mege, D
AU  - Mege D
FAU - Pascussi, J M
AU  - Pascussi JM
FAU - Dufour, E
AU  - Dufour E
FAU - Blondel, A
AU  - Blondel A
FAU - Acuto, O
AU  - Acuto O
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Receptors, Antigen, T-Cell)
RN  - 42HK56048U (Tyrosine)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.2 (Lymphocyte Specific Protein Tyrosine Kinase p56(lck))
RN  - EC 2.7.10.2 (ZAP-70 Protein-Tyrosine Kinase)
RN  - EC 2.7.10.2 (ZAP70 protein, human)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Binding Sites
MH  - Catalysis
MH  - Humans
MH  - Jurkat Cells
MH  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck)/*metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Phenotype
MH  - Phosphorylation
MH  - Protein-Tyrosine Kinases/genetics/*metabolism
MH  - Receptors, Antigen, T-Cell/genetics/*metabolism
MH  - Structure-Activity Relationship
MH  - Tyrosine/*metabolism
MH  - Up-Regulation
MH  - ZAP-70 Protein-Tyrosine Kinase
MH  - *src Homology Domains
EDAT- 1999/05/13 00:00
MHDA- 1999/05/13 00:01
CRDT- 1999/05/13 00:00
PHST- 1999/05/13 00:00 [pubmed]
PHST- 1999/05/13 00:01 [medline]
PHST- 1999/05/13 00:00 [entrez]
AID - 10.1074/jbc.274.20.14229 [doi]
AID - S0021-9258(19)73295-5 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 May 14;274(20):14229-37. doi: 10.1074/jbc.274.20.14229.