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Investigation of the interaction between CREB-binding protein and STAT4/STAT6

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Abstract

Coactivator CBP (CREB-binding protein) has been implicated in the regulation of transcription for all signal transducer and activator of transcription factors (STATs); however, the mechanism remains unclear. Using yeast two-hybrid screening and immunoprecipitation techniques, we investigated the direct interaction of CBP with STAT4 and STAT6. The full-length CBP and five fragments of CBP (residues 1–436, 529–1200, 1–697, 967–1574 and 1678–2175) were constructed using pGBKT7 vectors, while STAT4, STAT6 and N-terminal deleted STAT4 were constructed using pGADT7 vectors. It was found that STAT4, but not STAT6, interacted directly with the 1678–2175 fragment of CBP containing the ZZ, TAZ2 and SID domain. The N-terminal of STAT4 plays an important role in this interaction since N-terminal deleted STAT4 failed to bind to any CBP fragment. The results were confirmed by immunoprecipitation using HA-tagged STAT4 or STAT6 and c-Myc tagged CBP. This work will contribute to our understanding of the mechanisms of Th cytokine imbalance.

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Abbreviations

AP-1:

Activator protein-1

CBP:

CREB binding protein

CREB:

Cyclic AMP response element binding protein

JAK:

Janus kinase

NF-AT:

Nuclear factor of activated T cells

PRR:

Positive regulatory region

STAT:

Signal transducer and activator of transcription

TAD:

Transactivation domain

TSS:

Transcriptional start site

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Acknowledgments

This project was supported in part by the National Natural Science Foundation of China (No. 30672268). We thank Dr. Wei Liu at the Department of Clinical Immunology, the Children’s Hospital of Chongqing Medical University for primer design. We would also like to thank Prof. Xiongzhong Ruan for providing the pCDNA3.0-CBP plasmid DNA.

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Authors and Affiliations

  1. Department of Respiratory Medicine, Chongqing Medical University, Chongqing, 400014, People’s Republic of China

    Mingxiang Zhang, Zhou Fu, Daiyin Tian, Enmei Liu & Jihong Dai

  2. Clinical Immunology Research Laboratory, Children’s Hospital, Chongqing Medical University, Chongqing, People’s Republic of China

    Mingxiang Zhang, Zhou Fu, Daiyin Tian, Enmei Liu, Jihong Dai & Lijia Wang

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  1. Mingxiang Zhang
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  2. Zhou Fu
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Correspondence to Zhou Fu.

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Zhang, M., Fu, Z., Tian, D. et al. Investigation of the interaction between CREB-binding protein and STAT4/STAT6. Mol Biol Rep 38, 4805–4811 (2011). https://doi.org/10.1007/s11033-010-0622-0

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  • DOI: https://doi.org/10.1007/s11033-010-0622-0

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