ABCMdb

ABCG2 p.Thr362Ala

None has been submitted yet.

Publications

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

3 Knocking-down Pim-1L expression in the drug-resistant prostate cancer cells abolished multimer formation of endogenous BCRP and resensitized the resistant cells to chemotherapeutic drugs suggesting that BCRP phosphorylation induced byPim-1Lwasessentialforitsfunctionality.Thisisfurthercorrob- orated by our finding that the plasma membrane localization and drug-resistant activity of BCRP were compromised by T362A mutation. Login to 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). Login to comment
99 The stained slides of BCRP mutant T362A were examined by using an inverted fluorescence microscope under a ϫ60 oil-immersion objective, and the co-localization of BCRP with Pim-1L was scanned with a laser confocal system. Login to comment
150 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). Login to comment
152 As shown in Fig. 3D, this antibody recognized the wild-type BCRP phosphorylated by Pim-1L but not the unphosphorylytable T362A mutant. Login to comment
157 To test whether phosphorylation of Thr-362 of BCRP is important for BCRP-mediated drug resistance, we infected LNCaP cells with the lentivirus encoding the HA-tagged BCRP or the T362A mutant and then examined whether LNCaP cells were protected by the overexpression of BCRP or its mutant from apoptosis induced by chemotherapeutic drugs MX, TPT, and DTX. Login to comment
158 As shown in Fig. 4A, the wild-type BCRP significantly increased the survival of LNCaP cells in response to these drugs, but the T362A mutant failed to do so, suggesting that the integrity of Thr-362 is essential for the functionality of BCRP. Login to comment
163 A, the effects of BCRP phosphorylation site mutant T362A on drug-resistant activity. Login to comment
171 To test whether Thr-362 phosphorylation plays a role for BCRP membrane localization, we infected LNCaP cells by the lentivirus encoding the wild-type BCRP or the T362A mutant. Login to comment
172 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). Login to comment
173 The retention of BCRP in the cytoplasmic compartment caused by the T362A mutation was further confirmed by fractionation experiments (Fig. 5B). Login to comment
175 We therefore examined the effects of T362A mutation on BCRP dimerization. Login to comment
176 We co-transfected the wild-type HA-tagged BCRP with the Myc-tagged BCRP or its T362A mutant into 293T cells. Login to comment
178 As shown in Fig. 5C, the wild-type Myc-BCRP was efficiently associated with the wild-type HA-BCRP but such interaction was disrupted by the T362A mutation. Login to comment
190 Effects of T362A mutation on the subcellular distribution and multimerization of BCRP in prostate cancer cells. Login to comment
191 A, T362A mutant predominately localizes in the intracellular compartment. Login to comment
192 LNCaP cells were treated with the lentivirus encoding the wild-type BCRP, T362A, or T362D mutant. Login to comment
197 LNCaP cells were treated with the lentivirus encoding the wild-type BCRP or T362A mutant, or the vector control. Login to comment
201 C, T362A mutation disrupts the dimerization of BCRP. Login to comment
100 The stained slides of BCRP mutant T362A were examined by using an inverted fluorescence microscope under a ϫ60 oil-immersion objective, and the co-localization of BCRP with Pim-1L was scanned with a laser confocal system. Login to comment
151 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). Login to comment
153 As shown in Fig. 3D, this antibody recognized the wild-type BCRP phosphorylated by Pim-1L but not the unphosphorylytable T362A mutant. Login to comment
159 As shown in Fig. 4A, the wild-type BCRP significantly increased the survival of LNCaP cells in response to these drugs, but the T362A mutant failed to do so, suggesting that the integrity of Thr-362 is essential for the functionality of BCRP. Login to comment
164 A, the effects of BCRP phosphorylation site mutant T362A on drug-resistant activity. Login to comment
174 The retention of BCRP in the cytoplasmic compartment caused by the T362A mutation was further confirmed by fractionation experiments (Fig. 5B). Login to comment
177 We co-transfected the wild-type HA-tagged BCRP with the Myc-tagged BCRP or its T362A mutant into 293T cells. Login to comment
179 As shown in Fig. 5C, the wild-type Myc-BCRP was efficiently associated with the wild-type HA-BCRP but such interaction was disrupted by the T362A mutation. Login to comment
193 LNCaP cells were treated with the lentivirus encoding the wild-type BCRP, T362A, or T362D mutant. Login to comment
198 LNCaP cells were treated with the lentivirus encoding the wild-type BCRP or T362A mutant, or the vector control. Login to comment
202 C, T362A mutation disrupts the dimerization of BCRP. Login to comment

PMID: 23261525 [PubMed] Natarajan K et al: "The Pim kinase inhibitor SGI-1776 decreases cell surface expression of P-glycoprotein (ABCB1) and breast cancer resistance protein (ABCG2) and drug transport by Pim-1-dependent and -independent mechanisms."

No. Sentence Comment

22 Additionally, Pim-1 phosphorylates ABCG2 at threonine 362, knocking down Pim-1 abolishes ABCG2 multimer formation and ABCG2 T362A mutation decreases ABCG2 expression on the cell surface [18]. Login to comment

PMID: 26327810 [PubMed] Zhang W et al: "Insights into Chemoresistance of Prostate Cancer."

No. Sentence Comment

108 Moreover, the plasma membrane localization and drug-resistant activity of ABCG2 were compromised by T362A mutation, suggesting ABCG2 phosphorylation induced by Pim-1L is essential for ABCG2 functionality (78). Login to comment