Rac2 specificity in macrophage integrin signaling: potential role for Syk kinase.
Herein we report that, despite the similarity of Rac2 to Rac1 (92% amino acid identity), macrophages derived from Rac2-/- mice, which continue to express Rac1, display a marked defect in alphavbeta3/alphavbeta5 and alpha4beta1 integrin-directed migration measured on vitronectin and fibronectin fragments (FN-H296), respectively. In contrast, mouse embryo fibroblasts derived from the Rac2 knockout mice utilize Rac1 for migration via alphavbeta3/alphavbeta5 and alpha4beta1. The genetic reconstitution of bone marrow-derived macrophages (BMM) with Rac2 restores the integrin-dependent migration of Rac2-deficient macrophages on vitronectin (VN) and FN-H296. The levels of GTP-Rac2 generated upon specific integrin engagement in wild type macrophages parallels the phenotypic defect observed in Rac2-deficient macrophages; i.e. FN-H296, alpha4beta1 > VN, alphavbeta3/alphavbeta5 > FN-CH271, alpha5beta1 > intact FN. In a COS7 cell system, the expression of Syk kinase alone is sufficient to convert the alpha4beta1 migration response to Rac2 dependence. Therefore, we present the first evidence that the alpha4beta1 receptor in blood cells has evolved a Syk-Rac2 signaling axis to transmit signals required for integrin-directed migration suggesting that Syk kinase in part encodes myeloid Rac2 specificity in vivo.
Pradip
De
D
Section of Hematology/Oncology, Department of Pediatrics, Biochemistry and Molecular Biology, Herman B Wells Center for Pediatric Research, School of Medicine of Indiana University, 1044 W. Walnut Street, Indianapolis, IN 46202, USA.
Peng
Xiaodong
X
Durden
Donald L
DL
D000818:Animals; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002633:Chemotaxis; D004792:Enzyme Precursors / Q000502:physiology*; D005353:Fibronectins / Q000502:physiology; D020558:GTP Phosphohydrolases / Q000378:metabolism; D039041:Integrin alpha4beta1 / Q000502:physiology; D039302:Integrin alphaVbeta3 / Q000502:physiology; D016023:Integrins / Q000502:physiology; D047908:Intracellular Signaling Peptides and Proteins; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D019030:Receptors, Vitronectin / Q000502:physiology; D000072377:Syk Kinase; D019096:Vitronectin / Q000502:physiology; D020744:rac GTP-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D020830:rac1 GTP-Binding Protein / Q000032:analysis / Q000502:physiology; D000097766:RAC2 GTP-Binding Protein
D004792:Enzyme Precursors; D005353:Fibronectins; D020558:GTP Phosphohydrolases; D039041:Integrin alpha4beta1; D039302:Integrin alphaVbeta3; D016023:Integrins; D047908:Intracellular Signaling Peptides and Proteins; D011505:Protein-Tyrosine Kinases; D019030:Receptors, Vitronectin; D000072377:Syk Kinase; D019096:Vitronectin; D020744:rac GTP-Binding Proteins; D020830:rac1 GTP-Binding Protein; D000097766:RAC2 GTP-Binding Protein; C064660:integrin alphaVbeta5; C000606897:Syk protein, mouse