ABCG2 p.Thr362Asp
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PMID: 18056989
[PubMed]
Xie Y et al: "The 44-kDa Pim-1 kinase phosphorylates BCRP/ABCG2 and thereby promotes its multimerization and drug-resistant activity in human prostate cancer cells."
No.
Sentence
Comment
50
To generate the BCRP T362A or T362D mutant, the threonine residue at position 362 was mutated to alanine or aspartic acid via oligonucleotide-directed mutagenesis with the sense primers 5Ј-GAAGAAGATCGCAGTCTTCAAGG-3Ј or 5Ј-GAAG- AAGATCGACGTCTTCAAGG-3Ј, respectively and their complementary antisense primers by using the QuikChange Mutagenesis kit (Stratagene).
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ABCG2 p.Thr362Asp 18056989:50:30
status: VERIFIED150 Co-expression of BCRP and Pim-1L in 293T cells induced threonine phosphorylation of BCRP, and the substitution of Thr-362 with alanine (T362A) or the phosphorylation-mimicking aspartate (T362D) completely abolished threonine phosphorylation of BCRP, suggesting that Thr-362 may indeed be phosphorylated by Pim-1L in vivo (Fig. 3C).
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ABCG2 p.Thr362Asp 18056989:150:187
status: VERIFIED160 In addition, the BCRP drug-resistant activity was significantly reduced when endogenous Pim-1L expression was knocked down by the siRNA, whereas the phosphorylation-mimicking mutant T362D remained active independent of Pim-1L (Fig. 4B).
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ABCG2 p.Thr362Asp 18056989:160:182
status: VERIFIED166 *, p Ͻ 0.01 compared with the vector control. B, T362D mutant-mediated drug resistance is independent of Pim-1L.
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ABCG2 p.Thr362Asp 18056989:166:55
status: VERIFIED172 Immunofluorescence microscopy revealed that the wild-type BCRP and the phosphorylation- mimmicking mutant T362D were predominantly localized on plasma membrane, whereas the T362A mutant was mainly localized in cytoplasmic compartment (Fig. 5A).
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ABCG2 p.Thr362Asp 18056989:172:106
status: VERIFIED173 The retention of BCRP in the cytoplasmic compartment caused by the T362A mutation was further confirmed by fractionation experiments (Fig. 5B).
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ABCG2 p.Thr362Asp 18056989:173:106
status: NEW192 LNCaP cells were treated with the lentivirus encoding the wild-type BCRP, T362A, or T362D mutant.
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ABCG2 p.Thr362Asp 18056989:192:84
status: VERIFIED151 Co-expression of BCRP and Pim-1L in 293T cells induced threonine phosphorylation of BCRP, and the substitution of Thr-362 with alanine (T362A) or the phosphorylation-mimicking aspartate (T362D) completely abolished threonine phosphorylation of BCRP, suggesting that Thr-362 may indeed be phosphorylated by Pim-1L in vivo (Fig. 3C).
X
ABCG2 p.Thr362Asp 18056989:151:187
status: NEW161 In addition, the BCRP drug-resistant activity was significantly reduced when endogenous Pim-1L expression was knocked down by the siRNA, whereas the phosphorylation-mimicking mutant T362D remained active independent of Pim-1L (Fig. 4B).
X
ABCG2 p.Thr362Asp 18056989:161:182
status: NEW167 *, p Ͻ 0.01 compared with the vector control. B, T362D mutant-mediated drug resistance is independent of Pim-1L.
X
ABCG2 p.Thr362Asp 18056989:167:55
status: NEW193 LNCaP cells were treated with the lentivirus encoding the wild-type BCRP, T362A, or T362D mutant.
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ABCG2 p.Thr362Asp 18056989:193:84
status: NEW