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Two-dimensional NMR studies of d(GGTTAATGCGGT).d(ACCGCATTAACC) complexed with the minor groove binding drug SN-6999.
Chen SM, Leupin W, Rance M, Chazin WJ
(1992) Biochemistry 31: 4406-13
MeSH Terms: Antineoplastic Agents, Base Sequence, Binding Sites, Macromolecular Substances, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Nucleic Acid Conformation, Nucleic Acid Heteroduplexes, Oligodeoxyribonucleotides, Quinolinium Compounds, Structure-Activity Relationship
Show Abstract · Added December 10, 2013
The dodecadeoxynucleotide duplex d(GGTTAATGCGGT).d(ACCGCATTAACC) and its 1:1 complex with the minor groove binding drug SN-6999 have been prepared and studied by two-dimensional 1H nuclear magnetic resonance spectroscopy. Complete sequence-specific assignments have been obtained for the free duplex by standard methods. The line widths of the resonances in the complex are greater than those observed for the free duplex, which complicates the assignment process. Extensive use of two-quantum spectroscopy was required to determine the scalar correlations for identifying all of the base proton and most of the 1'H-2'H-2''H spin subsystems for the complex. This permitted unambiguous sequence-specific resonance assignments for the complex, which provides the necessary background for a detailed comparison of the structure of the duplex, with and without bound drug. A series of intermolecular NOEs between drug and DNA were identified, providing sufficient structural constraints to position the drug in the minor groove of the duplex. However, the combination of NOEs observed can only be rationalized by a model wherein the drug binds in the minor groove of the DNA in both orientations relative to the long helix axis and exchanges rapidly between the two orientations. The drug binds primarily in the segment of five consecutive dA-dT base pairs d(T3T4A5A6T7).d(A18T19T20A21A22), but surprisingly strong interactions are found to extend one residue in the 3' direction along each strand to G8 and C23. The observation of intermolecular contacts to residues neighboring the AT-rich region demonstrates that the stabilization of the bis(quaternary ammonium) heterocycle family of AT-specific, minor groove binding drugs is not based exclusively on interactions with dA-dT base pairs.
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11 MeSH Terms
Selection of drug-resistant bone marrow cells in vivo after retroviral transfer of human MDR1.
Sorrentino BP, Brandt SJ, Bodine D, Gottesman M, Pastan I, Cline A, Nienhuis AW
(1992) Science 257: 99-103
MeSH Terms: Alkaloids, Antineoplastic Agents, Phytogenic, Base Sequence, Bone Marrow, Bone Marrow Transplantation, DNA, Drug Resistance, Erythrocytes, Genetic Vectors, Hematopoietic Stem Cells, Humans, Molecular Sequence Data, Oligodeoxyribonucleotides, Paclitaxel, Polymerase Chain Reaction, Proviruses, Retroviridae, Transfection
Show Abstract · Added March 5, 2014
Experiments were performed to determine if retroviral-mediated transfer of the human multidrug resistance 1 gene (MDR1) into murine bone marrow cells would confer drug resistance to the cells and whether the MDR1 gene could be used as a dominant selectable marker in vivo. When mice transplanted with bone marrow cells containing a transferred MDR1 gene were treated with the cytotoxic drug taxol, a substantial enrichment for transduced bone marrow cells was observed. This demonstration of positive selection establishes the ability to amplify clones of transduced hematopoietic cells in vivo and suggests possible applications in human therapy.
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18 MeSH Terms