title: CK1delta modulates the transcriptional activity of ERalpha via AIB1 in an estrogen-dependent manner and regulates ERalpha-AIB1 interactions.
abstract: Oncogenesis in breast cancer often requires the overexpression of the nuclear receptor coactivator AIB1/SRC-3 acting in conjunction with estrogen receptor-alpha (ERalpha). Phosphorylation of both ERalpha and AIB1 has been shown to have profound effects on their functions. In addition, proteasome-mediated degradation plays a major role by regulating their stability and activity. CK1delta, a member of the ubiquitous casein kinase-1 family, is implicated in the progression of breast cancer. In this study, we show that both ERalpha and AIB1 are substrates for CK1delta in vitro, and identify a novel AIB1 phosphorylation site (S601) targeted by CK1delta, significant for the co-activator function of AIB1. CK1delta is able to interact with ERalpha and AIB1 in vivo, while overexpression of CK1delta in breast cancer cells results in an increased association of ERalpha with AIB1 as confirmed by co-immunoprecipitation assays from cell lysates. Using an siRNA-based approach, luciferase reporter assays and qRT-PCR, we observe that silencing of CK1delta leads to reduced ERalpha transcriptional activity, despite increased ERalpha levels, similarly to proteasome inhibition. We provide evidence that AIB1 protein levels are reduced by CK1delta silencing, in an estradiol-dependent manner; such destabilization can be inhibited by pre-treatment with the proteasome inhibitor MG132. We propose that differing activities adopted by ERalpha and AIB1 as a consequence of their interactions with and phosphorylation by CK1delta, particularly AIB1 stabilization, influence the transcriptional activity of ERalpha, and therefore have a role in breast cancer development.
authors.affiliation: Department of Medical Oncology, Imperial College London, Hammersmith Hospital Campus, London, UK.
authors.forename: Georgios
authors.initials: G
authors.lastname: Giamas
authors.forename: Leandro
authors.initials: L
authors.lastname: Castellano
authors.forename: Qin
authors.initials: Q
authors.lastname: Feng
authors.forename: Uwe
authors.initials: U
authors.lastname: Knippschild
authors.forename: Jimmy
authors.initials: J
authors.lastname: Jacob
authors.forename: Ross S
authors.initials: RS
authors.lastname: Thomas
authors.forename: R Charles
authors.initials: RC
authors.lastname: Coombes
authors.forename: Carolyn L
authors.initials: CL
authors.lastname: Smith
authors.forename: Long R
authors.initials: LR
authors.lastname: Jiao
authors.forename: Justin
authors.initials: J
authors.lastname: Stebbing