title: Inhibition of PRL-2·CNNM3 Protein Complex Formation Decreases Breast Cancer Proliferation and Tumor Growth.
abstract: The oncogenic phosphatase of regenerating liver 2 (PRL-2) has been shown to regulate intracellular magnesium levels by forming a complex through an extended amino acid loop present in the Bateman module of the CNNM3 magnesium transporter. Here we identified highly conserved residues located on this amino acid loop critical for the binding with PRL-2. A single point mutation (D426A) of one of those critical amino acids was found to completely disrupt PRL-2·human Cyclin M 3 (CNNM3) complex formation. Whole-cell voltage clamping revealed that expression of CNNM3 influenced the surface current, whereas overexpression of the binding mutant had no effect, indicating that the binding of PRL-2 to CNNM3 is important for the activity of the complex. Interestingly, overexpression of the CNNM3 D426A-binding mutant in cancer cells decreased their ability to proliferate under magnesium-deprived situations and under anchorage-independent growth conditions, demonstrating a PRL-2·CNNM3 complex-dependent oncogenic advantage in a more stringent environment. We further confirmed the importance of this complex in vivo using an orthotopic xenograft breast cancer model. Finally, because molecular modeling showed that the Asp-426 side chain in CNNM3 buries into the catalytic cavity of PRL-2, we showed that a PRL inhibitor could abrogate complex formation, resulting in a decrease in proliferation of human breast cancer cells. In summary, we provide evidence that this fundamental regulatory aspect of PRL-2 in cancer cells could potentially lead to broadly applicable and innovative therapeutic avenues.
authors.affiliation: From the Rosalind and Morris Goodman Cancer Research Centre, Montréal, Québec H3A 1A3, Canada, the Departments of Biochemistry and.
authors.forename: Elie
authors.initials: E
authors.lastname: Kostantin
authors.affiliation: From the Rosalind and Morris Goodman Cancer Research Centre, Montréal, Québec H3A 1A3, Canada.
authors.forename: Serge
authors.initials: S
authors.lastname: Hardy
authors.affiliation: Physiology, McGill University, Montréal, Québec H3A 0G4, Canada.
authors.forename: William C
authors.initials: WC
authors.lastname: Valinsky
authors.affiliation: the Department of Physiology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6500 HB Nijmegen, The Netherlands, and.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kompatscher
authors.affiliation: the Department of Physiology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6500 HB Nijmegen, The Netherlands, and.
authors.forename: Jeroen H F
authors.initials: JH
authors.lastname: de Baaij
authors.affiliation: From the Rosalind and Morris Goodman Cancer Research Centre, Montréal, Québec H3A 1A3, Canada, the Departments of Biochemistry and.
authors.forename: Yevgen
authors.initials: Y
authors.lastname: Zolotarov
authors.affiliation: From the Rosalind and Morris Goodman Cancer Research Centre, Montréal, Québec H3A 1A3, Canada.
authors.forename: Melissa
authors.initials: M
authors.lastname: Landry
authors.affiliation: From the Rosalind and Morris Goodman Cancer Research Centre, Montréal, Québec H3A 1A3, Canada.
authors.forename: Noriko
authors.initials: N
authors.lastname: Uetani
authors.affiliation: the Structural Biology Unit, Center for Cooperative Research in Biosciences (CIC bioGUNE), Technology Park of Bizkaia, 48160 Derio, Bizkaia, Spain.
authors.forename: Luis Alfonso
authors.initials: LA
authors.lastname: Martínez-Cruz
authors.affiliation: the Department of Physiology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6500 HB Nijmegen, The Netherlands, and.
authors.forename: Joost G J
authors.initials: JG
authors.lastname: Hoenderop
authors.affiliation: Physiology, McGill University, Montréal, Québec H3A 0G4, Canada.
authors.forename: Alvin
authors.initials: A
authors.lastname: Shrier
authors.affiliation: From the Rosalind and Morris Goodman Cancer Research Centre, Montréal, Québec H3A 1A3, Canada, the Departments of Biochemistry and michel.tremblay@mcgill.ca.
authors.forename: Michel L
authors.initials: ML
authors.lastname: Tremblay
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D017124:Conserved Sequence; D016213:Cyclins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D005260:Female; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008958:Models, Molecular; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D054730:Protein Interaction Domains and Motifs / Q000187:drug effects; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D011728:Pyridones / Q000494:pharmacology; D014410:Tumor Stem Cell Assay; D023041:Xenograft Model Antitumor Assays
chemical_list: C000597250:CNNM3 protein, human; D016213:Cyclins; D050505:Mutant Proteins; D011728:Pyridones; C487561:PTP4A2 protein, human; D017027:Protein Tyrosine Phosphatases
keywords: Bateman module; CNNM3; PRL-2; cancer; cell proliferation; magnesium; patch clamp; phosphatase; thienopyridone