Biochimie最新文献

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Inside front cover-EDB 内侧前盖EDB
Biochimie Pub Date : 2023-10-17 DOI: 10.1016/S0300-9084(23)00260-2
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引用次数: 0
Probing telomeric-like G4 structures with full or partial 2′-deoxy-5-hydroxyuridine substitutions 用2′-脱氧-5-羟基尿苷完全或部分取代探测类端粒G4结构
Biochimie Pub Date : 2023-01-24 DOI: 10.1016/j.biochi.2023.01.009
Zoltán Szeltner , Györgyi Ferenc , Tünde Juhász , Zoltán Kupihár , Zoltán Váradi , Dávid Szüts , Lajos Kovács
{"title":"Probing telomeric-like G4 structures with full or partial 2′-deoxy-5-hydroxyuridine substitutions","authors":"Zoltán Szeltner ,&nbsp;Györgyi Ferenc ,&nbsp;Tünde Juhász ,&nbsp;Zoltán Kupihár ,&nbsp;Zoltán Váradi ,&nbsp;Dávid Szüts ,&nbsp;Lajos Kovács","doi":"10.1016/j.biochi.2023.01.009","DOIUrl":"https://doi.org/10.1016/j.biochi.2023.01.009","url":null,"abstract":"<div><p><span><span>Guanine<span> quadruplexes (G4s) are stable four-stranded secondary DNA structures held together by noncanonical G-G base tetrads. We synthesised the </span></span>nucleoside analogue 2′-deoxy-5-hydroxyuridine (H) and inserted its phosphoramidite into telomeric repeat-type model </span>oligonucleotides<span><span><span><span>. Full and partial substitutions were made, replacing all guanines in all the three tetrads of a three-tier G4 structure, or only in the putative upper, central, or lower tetrads. We characterised these modified structures using CD, UV absorbance spectroscopy, native gel studies, and a capture oligo-based G4 disruption kinetic assay. The strand separation activity of BLM helicase on these substituted structures was also investigated. Two of the partially H-substituted constructs adopted G4-like structures, but displayed lower thermal stabilities compared to unsubstituted G4. The construct modified in its central tetrad remained mostly denatured, but the possibility of a special structure for the fully replaced variant remained open. H substitutions did not interfere with the G4-resolving activity of BLM helicase, but its efficiency was highly influenced by construct topology and even more by the G4 ligand PhenDC3. Our results suggest that the H modification can be incorporated into G quadruplexes, but only at certain positions to maintain G4 stability. The destabilizing effect observed for 2′-deoxy-5-hydroxyuridine indicates that the </span>cytosine </span>deamination product 5-hydroxyuracil and its nucleoside counterpart in </span>RNA (5-hydroxyuridine), might also be destabilizing in cellular DNA and RNA quadruplexes. The kinetic assay employed in this study can be generally employed for a fast comparison of the stabilities of various G4s either in their free or ligand-bound states.</span></p></div>","PeriodicalId":93898,"journal":{"name":"Biochimie","volume":"214 ","pages":"Pages 33-44"},"PeriodicalIF":0.0,"publicationDate":"2023-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49670757","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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