BACKGROUND: ICln is a multifunctional protein involved in the generation of chloride currents activated during regulatory volume decrease (RVD) after cell swelling (ICl(swell)). Growth factor receptors play a key role in different cellular processes and epidermal growth factor (EGF) regulates swelling-activated chloride permeability. AIM: We set out to investigate if the EGF-induced upregulation of ICl(swell) could be explained by a rearrangement of ICln subcellular distribution and interaction with its molecular partners. METHODS: NIH-3T3 fibroblasts were serum-deprived for 24 hours and stimulated with EGF (40 ng/ml) for 30 minutes. ICl(swell) activation, ICln distribution and interaction with its molecular partner HSPC038 were assessed by whole cell patch clamp and fluorescence resonance energy transfer (FRET). RESULTS: EGF treatment significantly enhanced the direct molecular interaction between ICln and HSPC038 and also resulted in an increase of ICln and HSPC038 association with the plasma membrane. Importantly, these events are associated with a significant increase of ICl(swell). CONCLUSIONS: The present data indicate that EGF might exert its role in the modulation of volume-sensitive chloride currents in part through activation and translocation of ICln and HSPC038 to the plasma membrane.
BACKGROUND:ICln is a multifunctional protein involved in the generation of chloride currents activated during regulatory volume decrease (RVD) after cell swelling (ICl(swell)). Growth factor receptors play a key role in different cellular processes and epidermal growth factor (EGF) regulates swelling-activated chloride permeability. AIM: We set out to investigate if the EGF-induced upregulation of ICl(swell) could be explained by a rearrangement of ICln subcellular distribution and interaction with its molecular partners. METHODS: NIH-3T3 fibroblasts were serum-deprived for 24 hours and stimulated with EGF (40 ng/ml) for 30 minutes. ICl(swell) activation, ICln distribution and interaction with its molecular partner HSPC038 were assessed by whole cell patch clamp and fluorescence resonance energy transfer (FRET). RESULTS:EGF treatment significantly enhanced the direct molecular interaction between ICln and HSPC038 and also resulted in an increase of ICln and HSPC038 association with the plasma membrane. Importantly, these events are associated with a significant increase of ICl(swell). CONCLUSIONS: The present data indicate that EGF might exert its role in the modulation of volume-sensitive chloride currents in part through activation and translocation of ICln and HSPC038 to the plasma membrane.
Authors: Roberta Costa; Alessia Remigante; Davide A Civello; Emanuele Bernardinelli; Zoltán Szabó; Rossana Morabito; Angela Marino; Antonio Sarikas; Wolfgang Patsch; Markus Paulmichl; Tamás Janáky; Attila Miseta; Tamás Nagy; Silvia Dossena Journal: Front Cell Dev Biol Date: 2020-11-19