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多核苷酸置换反应:通过干扰素诱导进行检测

Polynucleotide displacement reactions: detection by interferon induction.

作者信息

De Clercq E, Torrence P F, Witkop B

出版信息

Biochemistry. 1976 Feb 24;15(4):717-24. doi: 10.1021/bi00649a001.

Abstract

A large variety of displacement reactions between homopolynucleotides and complexes thereof has been demonstrated by interferon induction data obtained in primary rabbit kidney cell cultures superinduced with metabolic inhibitors. The polymers involved in these helix-coil displacement studies were: poly(adenylic acid), poly(inosinic acid), poly(cytidylic acid), poly(uridylic acid), poly(ribothymidylic acid), polylaurusin, poly(7-deazaadenylic acid), poly(7-deazainosinic acid), poly(5-bromocytidylic acid), and poly(5-bromouridylic acid). As monitored by ultraviolet absorbance-temperature profiles, all displacement reactions were directed toward the formation of the helix with the higher thermal stability. Concomitantly, the resulting helix was invariably more active as interferon inducer than the reactant helix, except for some reactions in which poly(7-deazaadenylic acid) was involved. For the latter reactions both the reactant and resultant helices were inactive as interferon inducer. The interferon induction data revealed that all displacement reactions proceeded to completion within 1 h even at temperatures well below the Tm of the reactant helix. The helix-coil displacement reaction could also be monitored by sucrose velocity gradient analysis, and, as evidenced for poly(A)-2poly(I) + 2poly(C) leads to 2poly(I)-poly(C) + poly(A), readily occurred at the cellular level, presumably at the cell surface.

摘要

在经代谢抑制剂超诱导的原代兔肾细胞培养物中获得的干扰素诱导数据表明,同聚核苷酸及其复合物之间存在多种置换反应。这些螺旋-卷曲置换研究中涉及的聚合物有:聚腺苷酸、聚肌苷酸、聚胞苷酸、聚尿苷酸、聚核糖胸苷酸、聚月桂素、聚7-脱氮腺苷酸、聚7-脱氮肌苷酸、聚5-溴胞苷酸和聚5-溴尿苷酸。通过紫外吸收-温度曲线监测发现,所有置换反应均朝着形成热稳定性更高的螺旋方向进行。与此同时,除了一些涉及聚7-脱氮腺苷酸的反应外,生成的螺旋作为干扰素诱导剂总是比反应物螺旋更具活性。对于后一类反应,反应物螺旋和生成的螺旋作为干扰素诱导剂均无活性。干扰素诱导数据显示,即使在远低于反应物螺旋熔点(Tm)的温度下,所有置换反应在1小时内也能进行到底。螺旋-卷曲置换反应也可以通过蔗糖速度梯度分析进行监测,并且正如聚(A)-2聚(I)+2聚(C)生成2聚(I)-聚(C)+聚(A)所证明的那样,这种反应在细胞水平上很容易发生,大概是在细胞表面。

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