Human monocyte chemotactic protein-2: cDNA cloning and regulated expression of mRNA in mesenchymal cells.
Stimulated MG-63 osteosarcoma cells have been used as a source to purify and identify the monocyte chemokines MCP-1, MCP-2 and MCP-3. In comparison with MCP-1, the production yields of MCP-2 and MCP-3 in these cells are rather low and variable. Although the protein sequence of human MCP-2 is identified, its DNA sequence remains elusive. A degenerate primer set was used to isolate an MCP-2 gene fragment from the chemokine YAC contig on human chromosome 17. Based on the gene sequence of MCP-2, a unique primer set was synthesized and used to screen cDNA libraries for the presence of MCP-2 transcripts by PCR. The complete MCP-2 cDNA was cloned from a human bone marrow cDNA library and sequenced. The cDNA-derived protein sequence was identical to that of purified natural MCP-2, except for Gln46 which replaced Lys46. There seem thus to exist two MCP-2 allelic variants because at position 46 the codons of two residues (Lys46 and Gln46) were detected in individual genomes. As shown by Northern hybridization, the MCP-2 steady-state mRNA levels in normal diploid fibroblasts were increased by IL-1 beta, IFN-gamma and the double-stranded RNA poly rI:rC. RT-PCR analysis showed induction of MCP-2 mRNA in MG-63 cells by IFN-gamma and IL-1 beta. The regulated production of MCP-2 by tumor cells and normal mesenchymal cells is indicative of a role in neoplasia and inflammatory host responses.
Van Coillie
E
E
Rega Institute for Medical Research, University of Leuven, Laboratory of Molecular Immunology, Louvain, Belgium.
Froyen
G
G
Nomiyama
H
H
Miura
R
R
Fiten
P
P
Van Aelst
I
I
Van Damme
J
J
Opdenakker
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D054412:Chemokine CCL8; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008648:Mesoderm / Q000502:physiology*; D008969:Molecular Sequence Data; D018945:Monocyte Chemoattractant Proteins / Q000235:genetics*; D011070:Poly I-C / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
C515453:CCL8 protein, human; D054412:Chemokine CCL8; D018076:DNA, Complementary; D007375:Interleukin-1; D008070:Lipopolysaccharides; D018945:Monocyte Chemoattractant Proteins; D012333:RNA, Messenger; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate; D011070:Poly I-C