TY - JOUR
T1 - CSF-1 receptor structure/function in MacCsf1r-/-macrophages
T2 - Regulation of proliferation, differentiation, and morphology
AU - Yu, Wenfeng
AU - Chen, Jian
AU - Xiong, Ying
AU - Pixley, Fiona J.
AU - Dai, Xu Ming
AU - Yeung, Yee Guide
AU - Stanley, E. Richard
PY - 2008/9/1
Y1 - 2008/9/1
N2 - CSF-1 is the major regulator of tissue macrophage development and function. A GM-CSF-dependent, CSF-1 receptor (CSF-1R)-deficient F4/80hiMac- 1+Gr1-CD11c+ bone marrow macrophage (BMM) line (MacCsf1r-/-) was developed to study the roles of the eight intracellular CSF-1R tyrosines phosphorylated upon receptor activation. Retroviral expression of the wild-type CSF-1R rescued the CSF-1-induced survival, proliferation, differentiation, and morphological characteristics of primary BMM. Mutation of all eight tyrosines failed to rescue, whereas the individual Y → F mutants (544, 559, 697, 706, 721, 807, 921, 974) rescued these CSF-1-inducible phenotypes to varying degrees. The juxtamembrane domain Y559F and activation loop Y807F mutations severely compromised proliferation and differentiation, whereas Y706, Y721F, and Y974F mutations altered morphological responses, and Y706F increased differentiation. Despite their retention of significant in vitro tyrosine kinase activity, Y559F and Y807F mutants exhibited severely impaired in vivo receptor tyrosine phosphorylation, consistent with the existence of cellular mechanisms inhibiting CSF-1R tyrosine phosphorylation that are relieved by phosphorylation of these two sites. The MacCsf1r-/- macrophage line will facilitate genetic and proteomic approaches to CSF-1R structure/function studies in the major disease-related CSF-1R-expressing cell type.
AB - CSF-1 is the major regulator of tissue macrophage development and function. A GM-CSF-dependent, CSF-1 receptor (CSF-1R)-deficient F4/80hiMac- 1+Gr1-CD11c+ bone marrow macrophage (BMM) line (MacCsf1r-/-) was developed to study the roles of the eight intracellular CSF-1R tyrosines phosphorylated upon receptor activation. Retroviral expression of the wild-type CSF-1R rescued the CSF-1-induced survival, proliferation, differentiation, and morphological characteristics of primary BMM. Mutation of all eight tyrosines failed to rescue, whereas the individual Y → F mutants (544, 559, 697, 706, 721, 807, 921, 974) rescued these CSF-1-inducible phenotypes to varying degrees. The juxtamembrane domain Y559F and activation loop Y807F mutations severely compromised proliferation and differentiation, whereas Y706, Y721F, and Y974F mutations altered morphological responses, and Y706F increased differentiation. Despite their retention of significant in vitro tyrosine kinase activity, Y559F and Y807F mutants exhibited severely impaired in vivo receptor tyrosine phosphorylation, consistent with the existence of cellular mechanisms inhibiting CSF-1R tyrosine phosphorylation that are relieved by phosphorylation of these two sites. The MacCsf1r-/- macrophage line will facilitate genetic and proteomic approaches to CSF-1R structure/function studies in the major disease-related CSF-1R-expressing cell type.
KW - Hematopoietic growth factor
KW - Receptor tyrosine kinase mutations
KW - Tyrosine kinase
KW - Tyrosine phosphorylation
UR - http://www.scopus.com/inward/record.url?scp=50849115188&partnerID=8YFLogxK
U2 - 10.1189/jlb.0308171
DO - 10.1189/jlb.0308171
M3 - Article
C2 - 18519746
AN - SCOPUS:50849115188
SN - 0741-5400
VL - 84
SP - 852
EP - 863
JO - Journal of Leukocyte Biology
JF - Journal of Leukocyte Biology
IS - 3
ER -