title: Carbonic anhydrase IX interacts with bicarbonate transporters in lamellipodia and increases cell migration via its catalytic domain.
abstract: Carbonic anhydrase IX (CA IX) is a hypoxia-induced cell surface enzyme expressed in solid tumors, and functionally involved in acidification of extracellular pH and destabilization of intercellular contacts. Since both extracellular acidosis and reduced cell adhesion facilitate invasion and metastasis, we investigated the role of CA IX in cell migration, which promotes the metastatic cascade. As demonstrated here, ectopically expressed CA IX increases scattering, wound healing and transwell migration of MDCK cells, while an inactive CA IX variant lacking the catalytic domain (ΔCA) fails to do so. Correspondingly, hypoxic HeLa cells exhibit diminished migration upon inactivation of the endogenous CA IX either by forced expression of the dominant-negative ΔCA variant or by treatment with CA inhibitor, implying that the catalytic activity is indispensable for the CA IX function. Interestingly, CA IX improves cell migration both in the absence and presence of hepatocyte growth factor (HGF), an established inducer of epithelial-mesenchymal transition. On the other hand, HGF up-regulates CA IX transcription and triggers CA IX protein accumulation at the leading edge of lamellipodia. In these membrane regions CA IX co-localizes with sodium bicarbonate co-transporter (NBCe1) and anion exchanger 2 (AE2) that are both components of the migration apparatus and form bicarbonate transport metabolon with CA IX. Moreover, CA IX physically interacts with AE2 and NBCe1 in situ, as shown here for the first time. Thus, our findings suggest that CA IX actively contributes to cell migration via its ability to facilitate ion transport and pH control at protruding fronts of moving cells.
authors.affiliation: Department of Molecular Medicine, Institute of Virology, Slovak Academy of Sciences, 845 05 Bratislava, Slovak Republic.
authors.forename: Eliska
authors.initials: E
authors.lastname: Svastova
authors.forename: Wojciech
authors.initials: W
authors.lastname: Witarski
authors.forename: Lucia
authors.initials: L
authors.lastname: Csaderova
authors.forename: Ivan
authors.initials: I
authors.lastname: Kosik
authors.forename: Lucia
authors.initials: L
authors.lastname: Skvarkova
authors.forename: Alzbeta
authors.initials: A
authors.lastname: Hulikova
authors.forename: Miriam
authors.initials: M
authors.lastname: Zatovicova
authors.forename: Monika
authors.initials: M
authors.lastname: Barathova
authors.forename: Juraj
authors.initials: J
authors.lastname: Kopacek
authors.forename: Jaromir
authors.initials: J
authors.lastname: Pastorek
authors.forename: Silvia
authors.initials: S
authors.lastname: Pastorekova
mesh_terms: D000818:Animals; D027321:Anion Transport Proteins / Q000235:genetics / Q000378:metabolism*; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D017920:Antiporters / Q000235:genetics / Q000378:metabolism*; D001639:Bicarbonates / Q000378:metabolism; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases / Q000096:biosynthesis* / Q000235:genetics; D015687:Cell Hypoxia / Q000502:physiology; D002465:Cell Movement / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006367:HeLa Cells; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017136:Ion Transport / Q000502:physiology; D017434:Protein Structure, Tertiary; D011554:Pseudopodia / Q000235:genetics / Q000378:metabolism*; D064606:SLC4A Proteins; D027982:Sodium-Bicarbonate Symporters / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation / Q000502:physiology
chemical_list: D027321:Anion Transport Proteins; D000951:Antigens, Neoplasm; D017920:Antiporters; D001639:Bicarbonates; C066855:HGF protein, human; D064606:SLC4A Proteins; C498898:SLC4A4 protein, human; D027982:Sodium-Bicarbonate Symporters; D017228:Hepatocyte Growth Factor; C089226:CA9 protein, human; D000071231:Carbonic Anhydrase IX; D002256:Carbonic Anhydrases