Abstract
SRC (steroid receptor co-activator)-1 has been reported to interact with and to be an essential co-activator for several members of the STAT (signal transducer and activator of transcription) family, including STAT3, the major signal transducer of IL (interleukin)-6. We addressed the question of whether SRC-1 is crucial for IL-6- and STAT3-mediated physiological responses such as myeloma cell survival and acute-phase protein induction. In fact, silencing of SRC-1 by RNA interference rapidly induced apoptosis in IL-6-dependent INA-6 human myeloma cells, comparable with what was observed upon silencing of STAT3. Using chromatin immunoprecipitation at STAT3 target regions of various genes, however, we observed constitutive binding of SRC-1 that decreased when INA-6 cells were treated with IL-6. The same held true for STAT3 target genes analysed in HepG2 human hepatocellular carcinoma cells. SRC-1-knockdown studies demonstrated that STAT3-controlled promoters require neither SRC-1 nor the other p160 family members SRC-2 or SRC-3 in HepG2 cells. Furthermore, microarray expression profiling demonstrated that the responsiveness of IL-6 target genes is not affected by SRC-1 silencing. In contrast, co-activators of the CBP [CREB (cAMP-response element-binding protein)-binding protein]/p300 family proved functionally important for the transactivation potential of STAT3 and bound inducibly to STAT3 target regions. This recruitment did not depend on the presence of SRC-1. Altogether, this suggests that functional impairment of STAT3 is not involved in the induction of myeloma cell apoptosis by SRC-1 silencing. We therefore conclude that STAT3 transactivates its target genes by the recruitment of CBP/p300 co-activators and that this process generally does not require the contribution of SRC-1.
- acute-phase reaction
- interleukin-6 (IL-6)
- signal transducer and activator of transcription 3 (STAT3)
- steroid receptor co-activator-1 (SRC-1)
- transcription control
Abbreviations: ACT, α1-antichymotrypsin; ChIP, chromatin immunoprecipitation; CREB, cAMP-response element-binding protein; CBP, CREB-binding protein; Crif1, CR6-interacting factor 1; DMEM, Dulbecco's modified minimal essential medium; EGFP, enhanced green fluorescent protein; ERK, extracellular-signal-regulated kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; gp, glycoprotein; GRIP-1, glutamate receptor-interacting protein 1; HEK, human embryonic kidney; IL, interleukin; IκBα, inhibitor of nuclear factor κB α; NCoA, nuclear receptor co-activator; NF-κB, nuclear factor κB; p/CAF, p300/CBP-associated factor; RNAi, RNA interference; shRNA, small hairpin RNA; SOCS3, suppressor of cytokine signalling 3; SRC, steroid receptor co-activator; STAT, signal transducer and activator of transcription; TRAP220, thyroid hormone receptor-associated polypeptide 220
- © The Authors Journal compilation © 2009 Biochemical Society