open access publication

Article, 2023

Dynamic localization of the Na+-HCO3− co-transporter NBCn1 to the plasma membrane, centrosomes, spindle and primary cilia

Journal of Cell Science, ISSN 1477-9137, 0021-9533, Volume 136, 7, 10.1242/jcs.260687

Contributors

Severin, Marc 0000-0003-4293-8059 [1] Pedersen, Emma Lind [1] Borre, Magnus Thane 0000-0002-9805-0019 [1] Axholm, Ida [1] Christiansen, Frederik Bendix [1] Ponniah, Muthulakshmi [1] Czaplinska, Dominika [1] Larsen, Tanja [1] Pardo, Luis Angel [2] Pedersen, Stine Falsig 0000-0002-3044-7714 [1]

Affiliations

  1. [1] University of Copenhagen
    2. [NORA names: KU University of Copenhagen; University; Denmark; Europe, EU; Nordic; OECD];
  2. [2] Max Planck Institute for Multidisciplinary Sciences
    3. [NORA names: Germany; Europe, EU; OECD]

Abstract

Finely tuned regulation of transport protein localization is vital for epithelial function. The Na+-HCO3- co-transporter NBCn1 (also known as SLC4A7) is a key contributor to epithelial pH homeostasis, yet the regulation of its subcellular localization is not understood. Here, we show that a predicted N-terminal β-sheet and short C-terminal α-helical motif are essential for NBCn1 plasma membrane localization in epithelial cells. This localization was abolished by cell-cell contact disruption, and co-immunoprecipitation (co-IP) and proximity ligation (PLA) revealed NBCn1 interaction with E-cadherin and DLG1, linking it to adherens junctions and the Scribble complex. NBCn1 also interacted with RhoA and localized to lamellipodia and filopodia in migrating cells. Finally, analysis of native and GFP-tagged NBCn1 localization, subcellular fractionation, co-IP with Arl13B and CEP164, and PLA of NBCn1 and tubulin in mitotic spindles led to the surprising conclusion that NBCn1 additionally localizes to centrosomes and primary cilia in non-dividing, polarized epithelial cells, and to the spindle, centrosomes and midbodies during mitosis. We propose that NBCn1 traffics between lateral junctions, the leading edge and cell division machinery in Rab11 endosomes, adding new insight to the role of NBCn1 in cell cycle progression.

Keywords

Arl13b, B sheets, CEP164, Co-IP, Dlg1, E-cadherin, NBCn1, Na+-HCO3, Rab11, Rab11 endosomes, RhoA, Scribble complex, adherens junctions, analysis, cell cycle progression, cell division machinery, cells, centrosome, cilia, co-immunoprecipitation, complex, contact disruption, cycle progression, disruption, division machinery, dynamic localization, edge, endosomes, epithelial cells, epithelial function, filopodia, finely tuned regulation, fines, fraction, function, homeostasis, interaction, junction, lamellipodia, lateral junctions, leading edge, ligation, localization, localized to lamellipodia, localizes to centrosomes, machinery, membrane, membrane localization, midbody, migrating cells, mitosis, mitotic spindle, motif, non-dividing, pH homeostasis, plasma, plasma membrane, plasma membrane localization, polarized epithelial cells, primary cilia, progression, protein localization, proximity, proximity ligation, regulation, scribbles, spindle, subcellular fractions, subcellular localization, transport, transport protein localization, tubulin

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