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[在聚[d(AT).d(AT)]模板存在下,大肠杆菌RNA聚合酶催化的多核苷酸合成反应中UTP的8-羟基-GTP底物特性展示]

[Display of 8-hydroxy-GTP substrate properties of UTP in the reaction of polynucleotide synthesis catalyzed by RNA-polymerase from Escherichia coli in the presence of poly[d(AT).d(AT)] template].

作者信息

Bruskov V I, Kuklina O V

出版信息

Mol Biol (Mosk). 1988 May-Jun;22(3):726-30.

PMID:3054496
Abstract

8-oxy-GTP was obtained via reaction of GTP with ascorbic acid and addition of hydrogen peroxide. 8-oxy-GTP is recognized and displays substrate properties of UTP on substitution of 8-oxy-GTP for UTP in polynucleotide synthesis catalyzed by E. coli RNA polymerase on a poly[d(A-T)].poly[d(A-T)] template. Such incorporation does not take place at equimolar quantities of GTP and 8-Br-GTP. The incorporation of 8-oxy-GTP instead of UTP, is 2.5-3 times higher upon replacement of Mg2+ by Mn2+ ions. The dinucleotide ApU serving as an initiator rises the incorporation level of 8-oxy-GTP both for Mg2+ and Mn2+ ions. 8-oxy-GTP slightly inhibits poly[r(A-U)] synthesis, but UTP strongly inhibits the incorporation of 8-oxy-GTP. [alpha-32P] 8-oxy-GTP is incorporated mainly instead of UTP, but it can be incorporated also during the substitution of 8-oxy-GTP for ATP.

摘要

8-氧代-GTP是通过GTP与抗坏血酸反应并添加过氧化氢获得的。在由大肠杆菌RNA聚合酶在聚[d(A-T)]·聚[d(A-T)]模板上催化的多核苷酸合成中,用8-氧代-GTP替代UTP时,8-氧代-GTP被识别并表现出UTP的底物特性。在等摩尔量的GTP和8-溴-GTP时不会发生这种掺入。用Mn2+离子替代Mg2+时,8-氧代-GTP替代UTP的掺入量高2.5至3倍。用作引发剂的二核苷酸ApU会提高Mg2+和Mn2+离子时8-氧代-GTP的掺入水平。8-氧代-GTP略微抑制聚[r(A-U)]的合成,但UTP强烈抑制8-氧代-GTP的掺入。[α-32P]8-氧代-GTP主要替代UTP掺入,但在8-氧代-GTP替代ATP时也可掺入。

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