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Effect of structural modifications on the activity of the leadzyme.

机译:结构修饰对铅酶活性的影响。

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The structure/function properties of functional groups in the leadzyme have been studied by assaying the activity of analog ribozymes generated by the systematic substitution of modified nucleotides in the internal loop region of the ribozyme. Guanosine analogs introduced at positions 4 and 7 occupied by guanosine in the wild-type molecule severely diminished cleavage. The substitution of deoxycytidine for cytidine at the cleavage site completely eliminated the activity of the leadzyme, as expected if the adjacent 2'-OH were the nucleophile in the cleavage reaction. On the other hand, substitution of an abasic nucleotide for adenosine at position 8 did not affect the activity of the ribozyme. An analysis of the activity of these analogs gives rise to the proposal of a triple-base pair motif implicating C1, G4, and G7.
机译:已经通过分析由核酶内部环区域中的修饰核苷酸的系统取代产生的类似核酶的活性,研究了先导酶中官能团的结构/功能特性。在野生型分子中由鸟苷占据的4和7位引入的鸟苷类似物严重减少了切割。如在裂解反应中相邻的2'-OH是亲核试剂所预期的,在裂解位点用脱氧胞苷取代胞苷完全消除了铅酶的活性。另一方面,用无碱基的核苷酸取代8位上的腺苷不会影响核酶的活性。对这些类似物活性的分析提出了涉及C1,G4和G7的三碱基对基序的提议。

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