CD2-mediated activation of the Tec-family tyrosine kinase ITK is controlled by proline-rich stretch-4 of the CD2 cytoplasmic tail.
Ligation of the CD2 co-stimulatory receptor on human T lymphocytes induces tyrosine phosphorylation and activation of the Tec-family tyrosine kinase, ITK. To examine whether any of several proline-rich (PR) stretches of the CD2 cytoplasmic tail are necessary for ITK activation we introduced wild-type and mutated versions of rat CD2, each missing at least one PR stretch of the tail, into human Jurkat T leukemia cells. The influence of cytoplasmic tail mutations was then studied following stimulation of transfectants with the rat CD2 mAb pair, OX54/OX55. As predicted, wild-type rat CD2 was able to activate ITK in Jurkat cells. In addition, a truncation mutant, lacking the most membrane-distal PR stretch, PR6, was able to activate ITK. By contrast, all other studied truncation mutants, each of which is missing at least PR4-PR6, were unable to induce ITK activation. Of deletion mutants, deletion of the membrane-proximal PR stretches, PR1-PR3, did not impair rat CD2-mediated ITK activation. However, additional deletion of PR4 from a tail missing PR1 and PR2, deletion of PR2 and PR4, and deletion of PR4 alone from rat CD2 abrogated an ability to activate ITK. Thus, these results identify PR4 as an element of the CD2 tail that is required for activation of ITK. Furthermore, we show that, unlike wild-type rat CD2, PR4-deleted rat CD2 is unable to induce IL-2 secretion from Jurkat cells. This is consistent with the view that PR4-mediated activation of ITK is important for downstream signaling events induced by CD2 co-stimulation.
Immunology Program, Sloan-Kettering Institute for Cancer Research, New York, NY 10021, USA.
P D
PD
King
A
A
Sadra
J M
JM
Teng
G M
GM
Bell
B
B
Dupont
D000818:Animals; D018801:CD2 Antigens / Q000235:genetics / Q000502:physiology*; D003593:Cytoplasm / Q000201:enzymology; D004789:Enzyme Activation; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis; D015458:Leukemia, T-Cell / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010455:Peptides / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D055232:Proline-Rich Protein Domains; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D013329:Structure-Activity Relationship; D014162:Transfection
D018801:CD2 Antigens; D007376:Interleukin-2; D010455:Peptides; D011505:Protein-Tyrosine Kinases; C080813:emt protein-tyrosine kinase