Open Access Research article

Ski-interacting protein (SKIP) interacts with androgen receptor in the nucleus and modulates androgen-dependent transcription

Daniel Abankwa16, Susan M Millard12, Nick Martel12, Catherine S Choong3, Miao Yang4, Lisa M Butler4, Grant Buchanan47, Wayne D Tilley4, Nobuhide Ueki5, Michael J Hayman5 and Gary M Leong12*

Author Affiliations

1 The University of Queensland, Obesity Research Centre, Institute for Molecular Bioscience, St.Lucia, Queensland, 4072, Australia

2 Department of Paediatric Endocrinology and Diabetes, Mater Children’s Hospital, South Brisbane, Queensland, 4010, Australia

3 Department of Paediatric Endocrinology and Diabetes, Princess Margaret Children’s Hospital, School of Pediatrics and Child Health, University of Western Australia, Subiaco, Western Australia, 6008, Australia

4 Dame Roma Mitchell Cancer Research Laboratories and Adelaide Prostate Cancer Research Centre, School of Medicine, University of Adelaide and Hanson Institute, Adelaide, South Australia, 5005, Australia

5 Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, 11794-5222, USA

6 Current addresses: Turku Centre for Biotechnology, Åbo Akademi University, Turku, Finland

7 Head, Cancer Biology Group, Freemasons Foundation Centre for Men’s Health, Basil Hetzel Institute for Translational Health Research, University of Adelaide, Adelaide, South Australia, 5005, Australia

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BMC Biochemistry 2013, 14:10  doi:10.1186/1471-2091-14-10

Published: 8 April 2013

Abstract

Background

The androgen receptor (AR) is a member of the nuclear receptor (NR) superfamily of ligand-inducible DNA transcription factors, and is the major mediator of male sexual development, prostate growth and the pathogenesis of prostate cancer. Cell and gene specific regulation by the AR is determined by availability of and interaction with sets of key accessory cofactors. Ski-interacting protein (SKIP; SNW1, NCOA62) is a cofactor shown to interact with several NRs and a diverse range of other transcription factors. Interestingly, SKIP as part of the spliceosome is thought to link mRNA splicing with transcription. SKIP has not been previously shown to interact with the AR.

Results

The aim of this study was to investigate whether SKIP interacts with the AR and modulates AR-dependent transcription. Here, we show by co-immunoprecipitation experiments that SKIP is in a complex with the AR. Moreover, SKIP increased 5α-dihydrotestosterone (DHT) induced N-terminal/C-terminal AR interaction from 12-fold to almost 300-fold in a two-hybrid assay, and enhanced AR ligand-independent AF-1 transactivation. SKIP augmented ligand- and AR-dependent transactivation in PC3 prostate cancer cells. Live-cell imaging revealed a fast (half-time=129 s) translocation of AR from the cytoplasm to the nucleus upon DHT-stimulation. Förster resonance energy transfer (FRET) experiments suggest a direct AR-SKIP interaction in the nucleus upon translocation.

Conclusions

Our results suggest that SKIP interacts with AR in the nucleus and enhances AR-dependent transactivation and N/C-interaction supporting a role for SKIP as an AR co-factor.