CD45 regulates Src family member kinase activity associated with macrophage integrin-mediated adhesion.
BACKGROUND
Adhesion of leukocytes to the extracellular matrix and to other cells is mediated by members of the integrin family of adhesion molecules. Src family kinases are activated upon integrin-mediated adhesion. In lymphocytes, CD45 is a leukocyte-specific transmembrane protein tyrosine phosphatase that activates Src family kinases associated with B-cell and T-cell antigen receptor signaling by constitutive dephosphorylation of the inhibitory carboxy-terminal tyrosine phosphorylation site. Here, we show that CD45 is also important in downregulating the kinase activity of Src family members during integrin-mediated adhesion in macrophages.


RESULTS
We found that CD45 colocalized with beta2 integrin and the Src family kinase p53/56(lyn) to adhesion sites in bone marrow-derived macrophages. Macrophages from CD45(-/-) mice were unable to maintain integrin-mediated adhesion. In adherent macrophages, absence of CD45 led to the hyperphosphorylation and hyperactivation of p56/59(hck) and p53/56(lyn), but not of p58(c-fgr). CD45 directly inactivated p59(hck) but not p56(lck) in transient transfection assays. Furthermore, coexpression of CD45 with p59(hck) or p56(lyn) containing a tyrosine to phenylalanine mutation at the carboxy-terminal negative regulatory site resulted in decreased tyrosine phosphorylation of the Src family member kinases due to dephosphorylation of the potentiating tyrosine phosphorylation site within the kinase domain.


CONCLUSIONS
Using primary bone marrow macrophages, these studies demonstrate that CD45 regulates Src family kinases and is required to maintain macrophage adhesion. CD45 decreases Src family kinase activity by dephosphorylating the tyrosine residue located within the kinase domain.
Roach
T
T
Division of Infectious Diseases, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Slater
S
S
Koval
M
M
White
L
L
Cahir McFarland
E D
ED
Okumura
M
M
Thomas
M
M
Brown
E
E
D000818:Animals; D018821:CD18 Antigens / Q000378:metabolism*; D002448:Cell Adhesion; D015536:Down-Regulation; D005786:Gene Expression Regulation; D016023:Integrins / Q000378:metabolism*; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D008817:Mice, Mutant Strains; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051572:Proto-Oncogene Proteins c-hck; D019061:src-Family Kinases / Q000378:metabolism*
D018821:CD18 Antigens; D016023:Integrins; D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; C496632:Hck protein, mouse; D051572:Proto-Oncogene Proteins c-hck; C060902:lyn protein-tyrosine kinase; C053102:proto-oncogene proteins c-fgr; D019061:src-Family Kinases; D017493:Leukocyte Common Antigens