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The linker region of breast cancer resistance protein ABCG2 is critical for coupling of ATP-dependent drug transport

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Abstract

The ATP-binding cassette (ABC) transporters of class G display a different domain organisation than P-glycoprotein/ABCB1 and bacterial homologues with a nucleotide-binding domain preceding the transmembrane domain. The linker region connecting these domains is unique and its function and structure cannot be predicted. Sequence analysis revealed that the human ABCG2 linker contains a LSGGE sequence, homologous to the canonical C-motif/ABC signature present in all ABC nucleotide-binding domains. Predictions of disorder and of secondary structures indicated that this C2-sequence was highly mobile and located between an α-helix and a loop similarly to the C-motif. Point mutations of the two first residues of the C2-sequence fully abolished the transport-coupled ATPase activity, and led to the complete loss of cell resistance to mitoxantrone. The interaction with potent, selective and non-competitive, ABCG2 inhibitors was also significantly altered upon mutation. These results suggest an important mechanistic role for the C2-sequence of the ABCG2 linker region in ATP binding and/or hydrolysis coupled to drug efflux.

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Abbreviations

ABC:

ATP-binding cassette

MDR:

Multidrug resistance

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Acknowledgments

Drs. A. Ahmed-Belkacem and C. Gauthier are acknowledged for their help in initiating the studies and performing some experiments, respectively. This work was supported by the CNRS and University Lyon 1 (UMR 5086), and the Ligue Nationale contre le Cancer (Equipe Labellisée Ligue 2014) to A.D.P. S.M. was recipient of fellowships from the Ligue de la Loire contre le Cancer, the Association pour la Recherche sur le Cancer and the Région Rhône-Alpes (Explora’Doc mobility program with S.E.B.). R.W.R., S.S., S.V.A. and S.E.B. were supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research (Z01 BC010030-17). The authors also thank research funding from OTKA K 111678 and the Bolyai Fellowship of the Hungarian Academy of Sciences to T.H.

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Authors and Affiliations

  1. Equipe Labellisée Ligue 2014, BMSSI, UMR 5086 CNRS, Université Lyon 1, IBCP, 69007, Lyon, France

    S. Macalou, G. Jabor Gozzi & A. Di Pietro

  2. Medical Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    R. W. Robey & S. E. Bates

  3. Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    S. Shukla & S. V. Ambudkar

  4. CelluloNet Biobank BB-0033-00072 Facility of UMS3444/US8/SFR Biosciences, IBCP, 69007, Lyon, France

    I. Grosjean

  5. MTA-SE Molecular Biophysics Research Group, Hungarian Academy of Sciences and Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary

    T. Hegedus

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  1. S. Macalou
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Correspondence to A. Di Pietro.

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Macalou, S., Robey, R.W., Jabor Gozzi, G. et al. The linker region of breast cancer resistance protein ABCG2 is critical for coupling of ATP-dependent drug transport. Cell. Mol. Life Sci. 73, 1927–1937 (2016). https://doi.org/10.1007/s00018-015-2118-5

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  • DOI: https://doi.org/10.1007/s00018-015-2118-5

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