Roles of the cytoplasmic domains of the alpha and beta subunits of human granulocyte-macrophage colony-stimulating factor receptor.
The high-affinity and functional granulocyte-macrophage colony-stimulating factor receptor (GMR) is composed of two distinct subunits, alpha and beta; and the cytoplasmic domain of the beta subunit is essential to transduce growth-promoting signals. In contrast to the beta subunit, the role of the alpha subunit is not well characterized. We examined the requirement of the cytoplasmic domain of the alpha subunit and its functional region by deletion analyses. We demonstrated that the cytoplasmic domain of the alpha subunit, especially 29 amino acids residues near the transmembrane domain, was absolutely required for various signaling events including activation of immediate early genes, induction of tyrosine phosphorylation of cellular proteins, and cell growth. We further analyzed the role of the cytoplasmic domain of each subunit by constructing chimeric subunits, designated alpha/beta and beta/alpha, by exchanging cytoplasmic domains of the alpha and beta subunits of human (h) GMR. Reconstituted high-affinity chimeric hGMRs, hGMR(alpha/beta,beta/alpha) and hGMR(alpha/beta,beta), transduced signals at levels similar to the wild type hGMR(alpha,beta) in Ba/F3 cells and in NIH3T3 cells. These observations indicate that the original configuration between the extracellular and the cytoplasmic domains of the hGMR(alpha,beta) subunits is not required and that hGMR(alpha/beta,beta) transduced signals through the cytoplasmic domain of the beta subunit in an oligomeric form, without involvement of the cytoplasmic domain of the alpha subunit. Therefore human granulocyte-macrophage colony-stimulating factor signals are mainly transduced through the beta subunit, and the cytoplasmic domain of the alpha subunit is likely to activate the beta subunit in the normal hGMR.
Muto
A
A
Department of Molecular and Developmental Biology, University of Tokyo, Japan.
Watanabe
S
S
Itoh
T
T
Miyajima
A
A
Yokota
T
T
Arai
K
K
D016475:3T3 Cells / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism / Q000648:ultrastructure; D005786:Gene Expression Regulation; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism
D046911:Macromolecular Substances; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; D011993:Recombinant Fusion Proteins; D014443:Tyrosine; D016178:Granulocyte-Macrophage Colony-Stimulating Factor