Alternative splicing of mRNA resulting in the deletion of a 94-bp transmembrane domain generates soluble interleukin-6 receptors in various cell lines and human peripheral blood mononuclear cells.
To detect transcripts encoding the interleukin-6 receptor (IL-6R) molecule lacking the transmembrane (TM) domain, in various cell lines and peripheral blood mononuclear cells (PBMC), we used the polymerase chain reaction (PCR) with primer pairs that flank the TM domain and which were selected to generate a 398-bp fragment. We detected 398-bp and 304-bp DNA molecules in the PCR products of the U1, J22HL60, MT-2, MT-4, U937 and HL60 cell lines and of PBMC isolated from several individuals. The sequencing analysis of both DNA molecules showed that a 94-bp region consisting of the TM domain of IL-6R was deleted in the 304-bp molecule. Moreover, we detected a soluble (s) IL-6R protein of 45 kDa in culture supernatants of the MT-2, MT-4 and U937 cell lines by radioimmunoprecipitation using specific antibodies against sIL-6R. Our results indicate that active deletion of the TM domain by alternative splicing of mRNA represents one mechanism for release of sIL-6R into the culture supernatants of cells, or into serum or urine.
Horiuchi et al. (Mon,) reported a other. Alternative splicing was evaluated on Detection of transcripts encoding IL-6R lacking the transmembrane domain. Alternative splicing of mRNA resulting in the deletion of a 94-bp transmembrane domain generates soluble interleukin-6 receptors in various cell lines and human peripheral blood mononuclear cells.