ABCMdb

ABCC3 p.Val1322Phe

None has been submitted yet.

Publications

PMID: 21799180 [PubMed] Henderson MJ et al: "ABCC multidrug transporters in childhood neuroblastoma: clinical and biological effects independent of cytotoxic drug Efflux."

No. Sentence Comment

69 For stable clones expressing ABCC3, BE(2)-C cells were transduced with retroviral pCMV14-3xFLAG-ABCC3 or the ATP-binding site mutant, pCMV14-3xFLAG-ABCC3-V1322F. Login to comment
70 The V1322F mutation was predicted to abolish transporter activity while still being localized in the plasma membrane and is based on studies involving the closely related transporter, ABCC6, in which a single amino acid change in a region N-terminal to the Walker A motif in the second ABC domain abolishes catalytic activity (20) while retaining correct membrane localization in mammalian cell lines (Andras Varadi, personal communication). Login to comment
75 Generation of ATP-binding site mutants was carried out accordingtotheQuikChangeLightningSite-DirectedMutagenesis Kit (Stratagene, La Jolla, CA) using forward primer 5'-ATC CTT GTG TTG AAT GAG GCC ACG GCA G-3' for ABCC1-D1454N (single mutant), forward primer 5'-ATC CTT GTG TTG CTT CTG GCC ACG GCA G-3' for ABCC1-DE1454/1455LL (double mutant) and forward primer 5'-AGG TGG GGA TCT TCG GCC GCA CTG G-3' for ABCC3-V1322F. Login to comment
281 ABCC3 transport activity is necessary for the observed effects on cell motility and clonogenicity, because the wound closure and colony-forming ability of cells expressing catalytically inactive ABCC3 (ABCC3-V1322F) were indistinguishable from cells transfected with empty vector (mean % of wound open ± 95% CI: ABCC3 C4, 5.4 ± 4.3 vs ABCC3 D1, 3.0 ± 2.4 vs empty vector, 1.5 ± 2.0, P = .34, one-way ANOVA, three independent experiments; Figure 6, C and mean numberof colonies ± 95% CI: ABCC3 C4, 91.1 ± 6.5 vs ABCC3 D1, 92.0 ± 7.4 vs empty vector, 90.9 ± 9.4, P = .98, one-way ANOVA, three independent experiments; Figure 6, D). Login to comment
283 A) Western blot analysis of ABCC3 protein expression following stable transduction of BE(2)-C cells with either empty vector, wild-type (wt) ABCC3 (clones A12, B12) or ABCC3 V1322F mutant (clones C4, D1) constructs. Login to comment

PMID: 26073088 [PubMed] Wiel C et al: "Multidrug resistance protein 3 loss promotes tumor formation by inducing senescence escape."

No. Sentence Comment

62 The pro-senescence effect of ABCC3 requires its transport activity ABCC3 is well known to act as a transporter of multiple macromolecules.17-19 To establish whether the pro-senescence effect of ABCC3 relies on its transporter activity, we generated a V1322F ABCC3 mutant. Login to comment
63 The V1322F mutation has been shown to inhibit the transport activity of ABCC proteins without altering its Figure 2. Login to comment
85 membrane localization.26 We infected HEC-TR cells with a control, ABCC3, or V1322F ABCC3 expression vector and checked for constitutive expression by immunoblotting (Figure 3a), RTqPCR (Figure 3b) and immunofluorescence (Supplementary Figure 2a). Login to comment
90 HEC-TR cells were infected with an empty control vector (ctrl), an ABCC3-encoding vector (ABCC3), or a vector encoding a transport-deficient ABCC3 mutant (ABCC3 V1322F) and neomycin selected. Login to comment
91 Generation of the ABCC3-V1322F mutant (ATP-binding site mutant) was carried out with and as recommended for the Quik Change Lightning Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA, USA). Login to comment
97 (c) MTT assay to measure etoposide efflux in HECs expressing an empty plasmid (ctrl) or a plasmid encoding wild-type ABCC3 or a mutated version thereof (V1322F). Login to comment
113 This confirms that ABCC3 V1322F has lost its transporter activity (Figure 3c). Login to comment
114 Strikingly, expression of the ABCC3 V1322F was unable to promote premature senescence: the results of the proliferation, SA-b2;-Gal activity, and IL8 expression assays showed no early effect after infection alone (Figure 3d, upper panel, 3e and f, left panel) and no effect at p6 in infected cells subjected to mild oncogenic stress (Figure 3d, lower panel, 3e and f, right panel). Login to comment