Molecular cloning of a second subunit of the receptor for human granulocyte-macrophage colony-stimulating factor (GM-CSF): reconstitution of a high-affinity GM-CSF receptor

Proc Natl Acad Sci U S A. 1990 Dec;87(24):9655-9. doi: 10.1073/pnas.87.24.9655.

Abstract

Using the mouse interleukin 3 (IL-3) receptor cDNA as a probe, we obtained a homologous cDNA (KH97) from a cDNA library of a human hemopoietic cell line, TF-1. The protein encoded by the KH97 cDNA has 56% amino acid sequence identity with the mouse IL-3 receptor and retains features common to the family of cytokine receptors. Fibroblasts transfected with the KH97 cDNA expressed a protein of 120 kDa but did not bind any human cytokines, including IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF). Interestingly, cotransfection of cDNAs for KH97 and the low-affinity human GM-CSF receptor in fibroblasts resulted in formation of a high-affinity receptor for GM-CSF. The dissociation rate of GM-CSF from the reconstituted high-affinity receptor was slower than that from the low-affinity site, whereas the association rate was unchanged. Cross-linking of 125I-labeled GM-CSF to fibroblasts cotransfected with both cDNAs revealed the same cross-linking patterns as in TF-1 cells--i.e., two major proteins of 80 and 120 kDa which correspond to the low-affinity GM-CSF receptor and the KH97 protein, respectively. These results indicate that the high-affinity GM-CSF receptor is composed of at least two components in a manner analogous to the IL-2 receptor. We therefore propose to designate the low-affinity GM-CSF receptor and the KH97 protein as the alpha and beta subunits of the GM-CSF receptor, respectively.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Line
  • Cloning, Molecular
  • Gene Library
  • Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
  • Humans
  • Kinetics
  • Mice
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Poly A / genetics
  • RNA / genetics
  • RNA, Messenger
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / genetics*
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / metabolism
  • Sequence Homology, Nucleic Acid
  • Transfection

Substances

  • Oligonucleotide Probes
  • RNA, Messenger
  • Receptors, Granulocyte-Macrophage Colony-Stimulating Factor
  • Poly A
  • RNA
  • Granulocyte-Macrophage Colony-Stimulating Factor

Associated data

  • GENBANK/M38275
  • GENBANK/M59941