4.5 Article

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

Journal

JOURNAL OF CELL SCIENCE
Volume 136, Issue 7, Pages -

Publisher

COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.260687

Keywords

KEY WORDS; Acid-base transport; Cell polarity; Cell-cell adhesion; Scribble complex; Primary cilium; Mitosis

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We found that the localization of NBCn1 in epithelial cells is regulated by a predicted N-terminal beta-sheet and short C-terminal alpha-helical motif. NBCn1 interacts with E-cadherin, DLG1, and RhoA, and localizes to adherens junctions, lamellipodia, filopodia, centrosomes, and primary cilia. Our findings suggest that NBCn1 plays a role in cell cycle progression by trafficking between lateral junctions, the leading edge, and cell division machinery.
Finely tuned regulation of transport protein localization is vital for 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 beta-sheet and short C-terminal alpha-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.

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