Low temperature restoring effect on F508del-CFTR misprocessing: A proteomic approach.
To gain insight into the proteins potentially involved in the low temperature-induced F508del-CFTR rescue process, we have explored by two-dimensional electrophoresis (2DE) the proteome of BHK cell lines expressing wt or F508del-CFTR, grown at 37 degrees C or 26 degrees C/24h or 26 degrees C/48h followed by 3h of metabolic labelling with [(35)S]-methionine. A set of 139 protein spots (yielding 125 mass spectrometry identifications) was identified as differentially expressed (p ANOVA<0.05) among the six phenotypic groups analysed. The data analysis suggests that the unfolded protein response (UPR) induction and some cell-metabolism repression are the major cold-shock responses that may generate a favourable cellular environment to promote F508del-CFTR rescue. Down-regulation of proteasome regulatory PA28 and/or COP9 signalosome subunit, both involved in CFTR degradation, could also be a relevant cold-shock-induced condition for F508de-CFTR rescue. Moreover, cold-shock may promote the reestablishment of some proteostasis imbalance associated with over-expression of F508del-CFTR. In BHK-F508del cells, the deregulation of RACK1, a protein described to be important for stable expression of CFTR in the plasma membrane, is partially repaired after low temperature treatment. Together these findings give new insights about F508del-CFTR rescue by low temperature treatment and the proteins involved could ultimately constitute potential therapeutic targets in CF disease.
Gomes-Alves
Patricia
P
Departamento de Genética, Instituto Nacional de Saúde Dr Ricardo Jorge, Lisboa, Portugal.
Neves
Sofia
S
Coelho
Ana V
AV
Penque
Deborah
D
D000222:Adaptation, Physiological; D000818:Animals; D002460:Cell Line; D003080:Cold Temperature; D006224:Cricetinae; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D016368:Frameshift Mutation; D019204:GTP-Binding Proteins; D005786:Gene Expression Regulation; D006801:Humans; D009363:Neoplasm Proteins; D040901:Proteomics / Q000379:methods*; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D013312:Stress, Physiological
D009363:Neoplasm Proteins; C419368:RACK1 protein, human; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019204:GTP-Binding Proteins