ABCMdb

PMID: 24058532

Venerando A, Franchin C, Cant N, Cozza G, Pagano MA, Tosoni K, Al-Zahrani A, Arrigoni G, Ford RC, Mehta A, Pinna LA
Detection of phospho-sites generated by protein kinase CK2 in CFTR: mechanistic aspects of Thr1471 phosphorylation.
PLoS One. 2013 Sep 18;8(9):e74232. doi: 10.1371/journal.pone.0074232. eCollection 2013., [PubMed]

Sentences

No. Mutations Sentence Comment

91 ABCC7 p.Thr1471Ala Cell Culture Baby hamster kidney cells (BHK) stably expressing human CFTR (hCFTR) either wild type (WT), or Phe508del, or WT T1471A were growth to reach 70-80% confluency [13]. Login to comment 147 ABCC7 p.Thr1471Ala Accordingly a hCFTR mutant no longer susceptible to such a phosphorylation (T1471A) was overexpressed in BHK cells as compared to hCFTR wild type. Login to comment 162 ABCC7 p.Thr1471Ala Once blockade is removed de novo synthesis of CFTR wild type is significantly stimulated if endogenous CK2 is inhibited by cell treatment with CX4945 (Panel A), a similar effect of CX4945 being also detectable if wild type CFTR is replaced by its T1471A mutant, which is no longer susceptible to phosphorylation at Thr1471 (Panel B). Login to comment 172 ABCC7 p.Thr1471Ala BHK cells expressing human CFTR wild type (A), T1471A (B) and Phe508del (C) were treated for 2 h with CHX to stop protein synthesis (compare first and second lanes). Login to comment 193 ABCC7 p.Thr1471Ala ABCC7 p.Thr1471Ala To get clues about the possible consequences of CFTR phosphorylation by CK2, advantage has been taken of the most selective CK2 inhibitor available to date, CX-4945 [43] and of a CFTR mutant in which Thr1471 has been replaced by alanine (T1471A) [13]. Login to comment 197 ABCC7 p.Thr1471Ala On the other hand the effect of CK2 on CFTR stability seems not to be mediated by Thr1471, at least in a wild type background, since a similar effect of CX4945 was observed with CFTR wild type and its T1471A mutant (Figure 5, compare Panels A and B). Login to comment 206 ABCC7 p.Thr1471Ala (TIF) Figure S4 BHK cells expressing WT, WT/T1471A or Phe508delCFTR were exposed to either cycloheximide (CHX, 100 mg/ml) or Brefeldin A (BFA, 200 ng/ml). Login to comment