Molecular cloning and functional expression of a new human CC-chemokine receptor gene

Biochemistry. 1996 Mar 19;35(11):3362-7. doi: 10.1021/bi952950g.

Abstract

The cloning of several receptors activated by either CC or CXC chemokines and belonging to the G protein-coupled family of receptors has been reported recently. In the present work, we describe the cloning of a human gene, named ChemR13, encoding a new CC-chemokine receptor. The gene encodes a protein of 352 amino acids with a calculated molecular mass of 40 600 Da and displaying a single potential site for N-linked glycosylation. Using a set of overlapping lambda clones, the genomic organisation of the locus was investigated, demonstrating that the ChemR13 gene is physically linked, and in the same orientation, as the CC-CKR2 gene that encodes a receptor for the monocyte chemoattractant protein-1 (MCP-1). A distance of 17.5 kb separates the two coding regions, which share 75% identity in nucleic acid and amino acid sequences. Human ChemR13 was functionally expressed in a stably transfected CHO-K1 cell line. Physiological responses to chemokines were monitored using a microphysiometer. Macrophage inflammatory protein 1 alpha (MIP-1 alpha) was the most potent agonist. MIP-1 beta and RANTES were also active at physiological concentrations. The other CC-chemokines, MCP-1, MCP-2 and MCP-3, as well as CXC-chemokines (IL-8, GRO alpha) had no effect. ChemR13 receptor transcripts were detected by Northern blotting in the promyeloblastic cell line KG-1A, suggesting a potential role in the control of granulocytic lineage proliferation or differentiation. ChemR13 is thus a new member of the growing family of chemokine receptors that mediate the recruitment of cells involved in immune and inflammatory processes. Being the fifth functionally identified receptor in his class, this new CC-chemokine receptor (CC-CKR) is tentatively designated CC-CKR5.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CHO Cells
  • Cloning, Molecular
  • Cricetinae
  • DNA Primers / chemistry
  • Gene Expression
  • Genes
  • Genetic Linkage
  • Humans
  • Molecular Sequence Data
  • Multigene Family
  • RNA, Messenger / genetics
  • Receptors, CCR5
  • Receptors, Cytokine / genetics*
  • Restriction Mapping
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • RNA, Messenger
  • Receptors, CCR5
  • Receptors, Cytokine

Associated data

  • GENBANK/U66285
  • GENBANK/X91492