[Construction of membrane-anchored interleukin-6 receptor beta subunit mutant and its effect on IL-6 signal transduction].
OBJECTIVE
To investigate whether the membrane-anchored gp130 mutant containing the second and the third fibronectin type III modules of the extracellular region could still transduce IL-6 signal.


METHODS
The membrane-anchored gp130 mutant cDNA was amplified by the overlap extension PCR and inserted into the mammalian expression vector pRc/RSV. The mutant was proved to be effectively expressed in SKO-007 cells and Jurkat cells by dot blot hybridization. The mutant's effects on IL-6 signaling in these two kinds of cells were detected by electrophoresis mobility shift assay (EMSA).


RESULTS
In SKO-007 cells, the mutant could enhance the activated APRF, but the effect was not as strong as that of wide-type gp130. IL-6 could not activate nuclear factors in Jurkat cells. After the expression vectors pRc/RSVgp130 and pRc/RSV were transfected into Jurkat cells, IL-6 could activate APRF, but could not do NF-IL6. Similar to the results in SKO-007 cells, the effects of the mutant was also weaker than that of wild-type gp130.


CONCLUSIONS
Although the mutant could still transduce IL-6 signal, it is not as efficiently as the wild-type gp130. It suggested that the Ig-like domain of gp130 also played an important role in IL-6 signal transduction.
Yang
Z
Z
Institute of Health and Environmental Medicine, Tianjin 300050, China.
Ni
C
C
Shen
B
B
Wang
J
J
Wang
Z
Z
D015703:Antigens, CD / Q000235:genetics* / Q000502:physiology; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000187:drug effects / Q000378:metabolism; D003001:Cloning, Molecular; D050822:Cytokine Receptor gp130; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D015850:Interleukin-6 / Q000494:pharmacology*; D019169:Jurkat Cells; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000502:physiology; D009154:Mutation; D050796:STAT3 Transcription Factor; D015398:Signal Transduction; D015534:Trans-Activators; D014162:Transfection; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D022782:CCAAT-Enhancer-Binding Protein-beta; D004268:DNA-Binding Proteins; C494199:IL6ST protein, human; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D015534:Trans-Activators; D050822:Cytokine Receptor gp130