Inhibition of protein-tyrosine kinase activity in intact cells by the aptameric action of oligodeoxynucleotides.

R C Bergan, E Kyle, Y Connell, L Neckers
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引用次数: 38

Abstract

Direct interaction of oligodeoxynucleotides (ODNs) with proteins represents one of the nonantisense-mediated effects of ODNs. Phosphorothioate-capped ODNs have been shown to inhibit directly the in vitro kinase activity of the chronic myelogenous leukemia-associated protein-tyrosine kinase p210bcr-abl. In this study we have determined the efficacy of this aptameric ODN in a cellular system using the K562 chronic myelogenous leukemia-derived cell line. Significant effects upon cellular phosphotyrosine content, as well as cellular growth in soft agar, are observed. These effects are sequence specific and are not mediated through changes in p210bcr-abl protein levels. Additional ODNs are described that also reduce cellular phosphotyrosine levels and inhibit growth in soft agar but do not inhibit p210bcr-abl kinase activity in vitro.

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寡脱氧核苷酸的自配体作用对完整细胞中蛋白酪氨酸激酶活性的抑制。
寡脱氧核苷酸与蛋白质的直接相互作用是寡脱氧核苷酸的非反义介导作用之一。硫代磷盖odn已被证明可以直接抑制慢性髓性白血病相关蛋白酪氨酸激酶p210bcr-abl的体外激酶活性。在这项研究中,我们利用K562慢性髓性白血病衍生细胞系确定了这种适配体ODN在细胞系统中的功效。在软琼脂中观察到对细胞磷酸酪氨酸含量以及细胞生长的显著影响。这些影响是序列特异性的,不通过p210bcr-abl蛋白水平的变化介导。其他odn也被描述为降低细胞磷酸酪氨酸水平并抑制软琼脂生长,但不抑制p210bcr-abl激酶活性。
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Detection of ribonuclease H-generated mRNA fragments in human leukemia cells following reversible membrane permeabilization in the presence of antisense oligodeoxynucleotides. (2'-5')-Oligo-3'-deoxynucleotides: selective binding to single-stranded RNA but not DNA. Inhibition of protein-tyrosine kinase activity in intact cells by the aptameric action of oligodeoxynucleotides. Helix-stabilizing compounds CC-1065 and U-71,184 bind to RNA-DNA and DNA-DNA duplexes containing modified internucleotide linkages and stabilize duplexes against thermal melting. Toward a broad-based antisense technology.
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