ATP13A3 is a major component of the enigmatic mammalian polyamine transport system

J Biol Chem. 2021 Jan-Jun;296:100182. doi: 10.1074/jbc.RA120.013908.

Norin Nabil Hamouda ¹, Chris Van den Haute ², Roeland Vanhoutte ³, Ragna Sannerud ⁴, Mujahid Azfar ¹, Rupert Mayer ⁵, Álvaro Cortés Calabuig ⁶, Johannes V Swinnen ⁷, Patrizia Agostinis ⁸, Veerle Baekelandt ⁹, Wim Annaert ⁴, Francis Impens ⁵, Steven H L Verhelst ¹⁰, Jan Eggermont ¹, Shaun Martin ¹, Peter Vangheluwe ¹¹

Affiliations

1 Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium.
2 Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Leuven, Belgium; Leuven Viral Vector Core, KU Leuven, Leuven, Belgium.
3 Laboratory of Chemical Biology, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium.
4 VIB-KU Leuven Laboratory of Membrane Trafficking, Department of Neurosciences, KU Leuven, Leuven, Belgium.
5 Department for Biomolecular Medicine, VIB Center for Medical Biotechnology, VIB Proteomics Core, Ghent University, Ghent, Belgium.
6 Genomics Core Leuven, KU Leuven, Leuven, Belgium.
7 Laboratory of Lipid Metabolism and Cancer, Department of Oncology, LKI - Leuven Cancer Institute, KU Leuven, Leuven, Belgium.
8 Laboratory of Cell Death Research & Therapy, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium; Department of Oncology, VIB-KU Leuven Center for Cancer Biology, KU Leuven, Leuven, Belgium.
9 Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Leuven, Belgium.
10 Laboratory of Chemical Biology, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium; Chemical Proteomics, Leibniz Institute for Analytical Sciences ISAS, Dortmund, Germany.
11 Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium. Electronic address: peter.vangheluwe@kuleuven.be.

PMID: 33310703 DOI: 10.1074/jbc.RA120.013908

Abstract

Polyamines, such as putrescine, spermidine, and spermine, are physiologically important polycations, but the transporters responsible for their uptake in mammalian cells remain poorly characterized. Here, we reveal a new component of the mammalian polyamine transport system using CHO-MG cells, a widely used model to study alternative polyamine uptake routes and characterize polyamine transport inhibitors for therapy. CHO-MG cells present polyamine uptake deficiency and resistance to a toxic polyamine biosynthesis inhibitor methylglyoxal bis-(guanylhydrazone) (MGBG), but the molecular defects responsible for these cellular characteristics remain unknown. By genome Sequencing of CHO-MG cells, we identified mutations in an unexplored gene, ATP13A3, and found disturbed mRNA and protein expression. ATP13A3 encodes for an orphan P5B-ATPase (ATP13A3), a P-type transport ATPase that represents a candidate polyamine transporter. Interestingly, ATP13A3 complemented the putrescine transport deficiency and MGBG resistance of CHO-MG cells, whereas its knockdown in WT cells induced a CHO-MG phenotype demonstrated as a decrease in putrescine uptake and MGBG sensitivity. Taken together, our findings identify ATP13A3, which has been previously genetically linked with pulmonary arterial hypertension, as a major component of the mammalian polyamine transport system that confers sensitivity to MGBG.

Keywords

ATP13A3; P-type ATPase; P5B-ATPase; polyamine; polyamine transport system; putrescine; transporter.

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