Melanoma growth stimulatory activity enhances the phosphorylation of the class II interleukin-8 receptor in non-hematopoietic cells.

Mueller SG, Schraw WP, Richmond A
J Biol Chem. 1994 269 (3): 1973-80

PMID: 8294449

The class II IL-8 receptor (IL-8R) binds both melanoma growth stimulatory activity (MGSA) and IL-8 with high affinity. Reverse transcriptase polymerase chain reaction demonstrates that the class II IL-8R mRNA, which has previously been detected only in cells of hematopoietic lineage, is also expressed in non-hematopoietic cell types shown to respond to MGSA or IL-8. To study the signaling mechanism by MGSA through the class II IL-8R in non-hematopoietic cells, this receptor was overexpressed in the 3ASubE human placental and the 293 human kidney cell lines. Membrane preparations of the class II IL-8R expressing 3ASubE transfectants exhibited a 2.3 +/- 0.2-fold increase in GTP gamma 35S binding, which was sensitive to pertussis toxin, in response to MGSA treatment (0.2 microM). This MGSA response was not observed in cells transfected with the parental expression vector. In vivo phosphorylation studies demonstrated that the class II IL-8R was basally phosphorylated in the untreated transfectants, and MGSA (5 nM) treatment markedly enhanced the phosphorylation of this receptor. The MGSA-induced receptor phosphorylation was both time and concentration dependent and could be mimicked by treatment with the calcium ionophore A23187. Phosphoamino acid analysis indicated that the MGSA-induced receptor phosphorylation was on serine residue(s), suggesting that a serine kinase is activated in response to MGSA binding to the class II IL-8R in non-hematopoietic cells.

MeSH Terms (31)

Base Sequence Cell Line Cell Membrane Cells, Cultured Chemokine CXCL1 Chemokines, CXC Chemotactic Factors Cloning, Molecular DNA Primers Female Growth Substances Guanosine 5'-O-(3-Thiotriphosphate) Humans Intercellular Signaling Peptides and Proteins Interleukin-8 Keratinocytes Kidney Kinetics Melanoma Molecular Sequence Data Neoplasm Proteins Open Reading Frames Phosphorylation Placenta Polymerase Chain Reaction Pregnancy Receptors, Interleukin Receptors, Interleukin-8A RNA, Messenger Transfection Tumor Cells, Cultured

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