Molecular cloning and functional expression of murine JE (monocyte chemoattractant protein 1) and murine macrophage inflammatory protein 1alpha receptors: evidence for two closely linked C-C chemokine receptors on chromosome 9.
We have isolated cDNA clones that encode two closely related, murine C-C chemokine receptors. Both receptors are members of the G-protein-coupled, seven-transmembrane domain family of receptors and are most closely related to the human monocyte chemoattractant protein 1 receptor. Expression of each of the receptors was detected in murine monocyte/macrophage cell lines, but not in nonhematopoietic lines. Expression of these receptors in Xenopus oocytes revealed that one receptor signaled in response to low nanomolar concentrations of murine JE, whereas the second receptor was activated by murine macrophage inflammatory protein (MIP) 1alpha and the human chemokines MIP-1beta and RANTES. Binding studies revealed high affinity binding of radiolabeled mJE to the mJE receptor and murine MIP-1alpha to the second receptor. Chromosomal localization indicated that the two receptor genes were clustered within 80 kilobases of each other on mouse chromosome 9. Creation of receptor chimeras suggested that the amino terminus was critically involved in mediating signal transduction and ligand specificity of the mJE receptor, but not the mMIP-1alpha receptor. The identification and cloning of two functional murine chemokine receptors provides important new tools for investigating the roles of these potent cytokines in vivo.
Boring
L
L
Gladstone Institute of Cardiovascular Disease, San Francisco, California 94141-9100, USA.
Gosling
J
J
Monteclaro
F S
FS
Lusis
A J
AJ
Tsou
C L
CL
Charo
I F
IF
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism; D015723:Gene Library; D008040:Genetic Linkage; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008263:Macrophage Migration-Inhibitory Factors / Q000378:metabolism; D051379:Mice / Q000235:genetics*; D008810:Mice, Inbred C57BL / Q000235:genetics; D008969:Molecular Sequence Data; D009115:Muridae / Q000235:genetics; D009865:Oocytes / Q000502:physiology; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D018121:Receptors, Cytokine / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D014162:Transfection
C515334:CCR2 protein, human; C515335:Ccr2 protein, mouse; D018932:Chemokine CCL2; D008263:Macrophage Migration-Inhibitory Factors; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D018121:Receptors, Cytokine; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C080633:macrophage inflammatory protein 1alpha receptor; D019204:GTP-Binding Proteins; D002118:Calcium