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C-terminal-binding protein interacting protein binds directly to adenovirus early region 1A through its N-terminal region and conserved region 3

Abstract

C-terminal-binding protein interacting protein (CtIP) was first isolated as a binding partner of C-terminal-binding protein (CtBP). It is considered to contribute to the transcriptional repression and cell cycle regulatory properties of the retinoblastoma (Rb) family of proteins and to have a role in the cellular response to DNA damage. Here, we have shown that CtIP is a novel target for the adenovirus oncoprotein early region 1A (AdE1A). AdE1A associates with CtIP in both Ad5E1-transformed cells and Ad5-infected cells and binds directly in glutathione-S-transferase pull-down assays. Two binding sites have been mapped on Ad5E1A – the N-terminal α-helical region (residues 1–30) and conserved region 3 (CR3) – the transcriptional activation domain. CtIP can bind AdE1A and CtBP independently, raising the possibility that ternary complexes exist in Ad-transformed and -infected cells. Significantly, reduction of CtIP expression with small interfering RNAs results in reduction of the ability of a Gal4 DNA-binding domain-CR3 construct to transactivate a Gal 4-responsive luciferase reporter and this effect is reversed by reduction of CtBP expression. Therefore, in this model, CtIP acts as a transcriptional co-activator of AdE1A when dissociated from CtBP, through the action of AdE1A. These data are consistent with observations that CtIP expression is induced by AdE1A during viral infection and that reduction of CtIP expression with RNA interference can retard virus replication. In addition, AdE1A causes disruption of the CtIP/Rb complex during viral infection by its interaction with CtIP, possibly contributing to transcriptional derepression.

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Acknowledgements

We are most grateful to Professor Richard Baer (Columbia University, USA) for the CtIP constructs and Dr Richard Lundblad (Salk Institute, USA) for the gift of His-tagged CtBP1. We thank Dr Vivien Mautner and Dr Thomas Dobner for gifts of antibodies. We also thank Dr Mautner for help with the quantitative PCR experiments. We thank Cancer Research UK, the Breast Cancer Campaign and the CIHR for generous funding and the Iranian government for a PhD studentship to MR.

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Correspondence to R J A Grand.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Bruton, R., Rasti, M., Mapp, K. et al. C-terminal-binding protein interacting protein binds directly to adenovirus early region 1A through its N-terminal region and conserved region 3. Oncogene 26, 7467–7479 (2007). https://doi.org/10.1038/sj.onc.1210551

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