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Identification of PLP2 and RAB5C as novel TPD52 binding partners through yeast two-hybrid screening

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Abstract

Tumor protein D52 (TPD52) is overexpressed in different cancers, but its molecular functions are poorly defined. A large, low-stringency yeast two-hybrid screen using full-length TPD52 bait identified known partners (TPD52, TPD52L1, TPD52L2, MAL2) and four other preys that reproducibly bound TPD52 and TPD52L1 baits (PLP2, RAB5C, GOLGA5, YIF1A). PLP2 and RAB5 interactions with TPD52 were confirmed in pull down assays, with interaction domain mapping experiments indicating that both proteins interact with a novel binding region of TPD52. This study provides insights into TPD52 functions, and ways to maximise the efficiency of low-stringency yeast two-hybrid screens.

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Acknowledgments

This work was supported by a project grant from the New South Wales Cancer Council (to JAB and BJS), a National Health and Medical Research Council of Australia postdoctoral fellowship (to YC), and Endeavour International- and International Postgraduate Research Scholarships (EIPRS and IPRS) (to HS), and by donations to the Children’s Cancer Research Unit of the Children’s Hospital at Westmead. The authors thank Dr Mona Shehata (CHW), Dr Erdahl Teber and A/Prof Jonathan Arthur (Children’s Medical Research Institute, Australia) for discussions, and Prof Philip D. Stahl (Washington University, USA) for ECFP-RAB5 expression plasmids.

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Authors and Affiliations

  1. Children’s Cancer Research Unit, Kids Research Institute, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia

    Hamideh Shahheydari, Sarah Frost, Yuyan Chen & Jennifer A. Byrne

  2. The University of Sydney Discipline of Paediatrics and Child Health, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia

    Hamideh Shahheydari, Sarah Frost, Yuyan Chen & Jennifer A. Byrne

  3. Department of Chemistry, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC, 3086, Australia

    Brian J. Smith

  4. Department of Nutritional Sciences, University of Wisconsin-Madison, Madison, WI, USA

    Guy E. Groblewski

Authors
  1. Hamideh Shahheydari
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  2. Sarah Frost
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  3. Brian J. Smith
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  4. Guy E. Groblewski
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  5. Yuyan Chen
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  6. Jennifer A. Byrne
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Correspondence to Jennifer A. Byrne.

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Shahheydari, H., Frost, S., Smith, B.J. et al. Identification of PLP2 and RAB5C as novel TPD52 binding partners through yeast two-hybrid screening. Mol Biol Rep 41, 4565–4572 (2014). https://doi.org/10.1007/s11033-014-3327-y

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  • DOI: https://doi.org/10.1007/s11033-014-3327-y

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