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.
Departamento de Genética, Instituto Nacional de Saúde Dr Ricardo Jorge, Lisboa, Portugal.
Patricia
P
Gomes-Alves
Sofia
S
Neves
Ana V
AV
Coelho
Deborah
D
Penque
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