ABCMdb

PMID: 26073088

Wiel C, Gras B, Vindrieux D, Warnier M, Gitenay D, Le Calve B, Ferrand M, Augert A, Bernard D
Multidrug resistance protein 3 loss promotes tumor formation by inducing senescence escape.
Oncogene. 2015 Jun 15. doi: 10.1038/onc.2015.218., [PubMed]

Sentences

No. Mutations Sentence Comment

62 ABCC3 p.Val1322Phe 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 ABCC3 p.Val1322Phe The V1322F mutation has been shown to inhibit the transport activity of ABCC proteins without altering its Figure 2. Login to comment 85 ABCC3 p.Val1322Phe 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 ABCC3 p.Val1322Phe 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 ABCC3 p.Val1322Phe 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 ABCC3 p.Val1322Phe (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 ABCC3 p.Val1322Phe This confirms that ABCC3 V1322F has lost its transporter activity (Figure 3c). Login to comment 114 ABCC3 p.Val1322Phe 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