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Molecular Pharmacology

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Abstract

Structure and expression of the messenger RNA encoding the murine multidrug resistance protein, an ATP-binding cassette transporter.

B D Stride, G Valdimarsson, J H Gerlach, G M Wilson, S P Cole and R G Deeley
Molecular Pharmacology June 1996, 49 (6) 962-971;
B D Stride
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G Valdimarsson
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J H Gerlach
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G M Wilson
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S P Cole
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R G Deeley
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Abstract

In vitro, overexpression of the human multidrug-resistance protein (MRP) causes a form of multidrug resistance similar to that conferred by P-glycoprotein, although the two proteins are only very distantly related. Studies with MRP-enriched membrane vesicles have demonstrated that the protein can bind and transport cysteinyl leukotrienes, as well as some other glutathione conjugates, with high affinity. In contrast, there is no direct evidence of the ability of MRP to bind or transport unmodified forms of the drugs to which it confers resistance. To facilitate studies of the physiological function(s) of MRP and its ability to cause multidrug resistance in vivo, we cloned and characterized the mRNA specifying its murine homolog. The murine MRP mRNA encodes a protein of 1528 amino acids that is 88% identical to human MRP. Although detectable by Northern blotting at variable levels in a wide range of tissues, in situ hybridization experiments revealed that MRP mRNA expression in some tissues is cell-type specific. High levels of the mRNA were detected in epithelia lining bronchi and bronchioles, as well as stage-specific expression in the seminiferous epithelium of the testes. Comparison of the predicted hydropathy profiles of human and murine MRP suggests a highly conserved membrane topology, the most distinctive feature of which is an extremely hydrophobic NH2-terminal region containing five or six potential transmembrane sequences. This structural feature is shared with the sulfonylurea receptor and the yeast cadmium factor 1 but is not present in members of the superfamily, such as the cystic fibrosis transmembrane conductance regulator and P-glycoproteins. Finally, we used overlapping cDNAs to construct an episomally replicating murine MRP expression vector that was stably transfected into HeLa cells. MRP-Transfected cell populations expressed markedly elevated levels of a 180-190-kDa protein that cross-reacted with a polyclonal antiserum raised against a peptide that is completely conserved in murine and human MRPs. The MRP transfectants also displayed increased resistance to vincristine (5-6-fold) and doxorubicin (< 2-fold).

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Molecular Pharmacology
Vol. 49, Issue 6
1 Jun 1996
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Abstract

Structure and expression of the messenger RNA encoding the murine multidrug resistance protein, an ATP-binding cassette transporter.

B D Stride, G Valdimarsson, J H Gerlach, G M Wilson, S P Cole and R G Deeley
Molecular Pharmacology June 1, 1996, 49 (6) 962-971;

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Abstract

Structure and expression of the messenger RNA encoding the murine multidrug resistance protein, an ATP-binding cassette transporter.

B D Stride, G Valdimarsson, J H Gerlach, G M Wilson, S P Cole and R G Deeley
Molecular Pharmacology June 1, 1996, 49 (6) 962-971;
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