A simple bypass assay for DNA polymerases shows hypermutating variants associated with cancer show mechanistic differences in vitro
A simple bypass assay for DNA polymerases shows hypermutating variants associated with cancer show mechanistic differences in vitro
复制标题
DNA 聚合酶的简单旁路检测显示与癌症相关的超突变变异在体外表现出机制差异
DOI：
10.1101/2022.01.10.475213
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Crevel G
中科院分区：
文献类型：
--
作者：
Crevel G
由DNA聚合酶造成的错误既会导致自然变异，也会在极端情况下导致基因组不稳定及其相关疾病。最近，聚合酶错误掺入疾病的重要性已经突出了癌症相关的聚合酶变异体的鉴定和认识到，这些变异体的一个亚组在肿瘤中具有超突变表型。我们已经开发了一种旁路试验，以快速确定聚合酶在体外错误掺入的趋势。我们已经使用该测定来比较野生型、核酸外切酶缺陷型和两种超突变DNA聚合酶e变体P286R和V411L的错误掺入。该测定清楚地区分了野生型、外切核酸酶死亡和P286R聚合酶的错误掺入率。然而，V411L聚合酶表现出与P286R不同的错误掺入特征，表明这些变体通过不同的机制引起超突变。使用该测定，相对于模板化的C核苷酸的错误掺入始终高于其他核苷酸，并且这主要导致C至T的转换。这与在POLE突变体肿瘤中常见的C至T转换的观察结果一致。
Errors made by DNA polymerases contribute to both natural variation and, in extreme cases, to genome instability and its associated diseases. Recently the importance of polymerase misincorporation in disease has been highlighted by the identification of cancer-associated polymerase variants and the recognition that a subgroup of these variants have a hypermutation phenotype in tumours. We have developed a bypass assay to rapidly determine the tendency of a polymerase to misincorporate in vitro. We have used the assay to compare misincorporation by wild-type, exonuclease defective and two hypermutating DNA polymerase e variants, P286R and V411L. The assay clearly distinguished between the misincorporation rates of wild type, exonuclease dead and P286R polymerases. However, the V411L polymerase showed different misincorporation characteristics to P286R, suggesting that these variants cause hypermutation by different mechanisms. Using this assay misincorporation opposite a templated C nucleotide was consistently higher than for other nucleotides, and this caused predominantly C to T transitions. This is consistent with the observation that C to T transitions are commonly seen in POLE mutant tumours.
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影响因子：
14.9
作者：
Herzog M;Alonso-Perez E;Salguero I;Warringer J;Adams DJ;Jackson SP;Puddu F
通讯作者：
Puddu F
影响因子：
7
作者：
Shinbrot E;Henninger EE;Weinhold N;Covington KR;Göksenin AY;Schultz N;Chao H;Doddapaneni H;Muzny DM;Gibbs RA;Sander C;Pursell ZF;Wheeler DA
通讯作者：
Wheeler DA
影响因子：
5.2
作者：
Pavlov YI;Zhuk AS;Stepchenkova EI
通讯作者：
Stepchenkova EI
影响因子：
16.6
作者：
Burgers PMJ;Kunkel TA
通讯作者：
Kunkel TA
影响因子：
16.6
作者：
Lancey，Claudia;Tehseen，Muhammad;De Biasio，Alfredo
通讯作者：
De Biasio，Alfredo