2-Substituted 2'-deoxyinosine 5'-triphosphates as substrates for polymerase synthesis of minor-groove-modified DNA and effects on restriction endonuclease cleavage

0520880 - ÚOCHB 2021 RIV GB eng J - Journal Article
Matyašovský, Ján - Hocek, Michal
2-Substituted 2'-deoxyinosine 5'-triphosphates as substrates for polymerase synthesis of minor-groove-modified DNA and effects on restriction endonuclease cleavage.
Organic & Biomolecular Chemistry. Roč. 18, č. 2 (2020), s. 255-262. ISSN 1477-0520. E-ISSN 1477-0539
R&D Projects: GA ČR(CZ) GA17-03419S; GA MŠMT(CZ) EF16_019/0000729
EU Projects: European Commission(XE) 642023 - ClickGene
Institutional support: RVO:61388963
Keywords : 2'-deoxyribonucleoside triphosphates * nucleoside triphosphates * primer extension
OECD category: Organic chemistry
Impact factor: 3.876, year: 2020
Method of publishing: Limited access
https://pubs.rsc.org/en/content/articlelanding/2020/OB/C9OB02502B#!divAbstract

Five 2-substituted 2'-deoxyinosine triphosphates (d (R) ITP) were synthesized and tested as substrates in enzymatic synthesis of minor-groove base-modified DNA. Only 2-methyl and 2-vinyl derivatives proved to be good substrates for Therminator DNA polymerase, whilst all other d (R) ITPs and other tested DNA polymerases did not give full length products in primer extension. The DNA containing 2-vinylhypoxanthine was then further modified through thiol-ene reactions with thiols. Cross-linking reaction between cysteine-containing minor-groove binding dodecapeptide and DNA proceeded thanks to the proximity effect between thiol and vinyl groups inside the minor groove. 2-Substituted dI (R) TPs and also previously prepared 2-substituted 2'-deoxyadenosine triphosphates (d (R) ATP) were then used for enzymatic synthesis of minor-groove modified DNA to study the effect of minor-groove modifications on cleavage of DNA by type II restriction endonucleases (REs). Although the REs should recognize the sequence through H-bonds in the major groove, some minor-groove modifications also had an inhibiting effect on the cleavage.
Permanent Link: http://hdl.handle.net/11104/0305542