Influence of major-groove chemical modifications of DNA on transcription by bacterial RNA polymerases

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F16%3A00461200" target="_blank" >RIV/61388971:_____/16:00461200 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388963:_____/16:00461200 RIV/00216208:11310/16:10323877 RIV/00216208:11320/16:10323877

Výsledek na webu

<a href="http://nar.oxfordjournals.org/content/44/7/3000.full" target="_blank" >http://nar.oxfordjournals.org/content/44/7/3000.full</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1093/nar/gkw171" target="_blank" >10.1093/nar/gkw171</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Influence of major-groove chemical modifications of DNA on transcription by bacterial RNA polymerases

Popis výsledku v původním jazyce

DNA templates containing a set of base modifications in the major groove (5-substituted pyrimidines or 7-substituted 7-deazapurines bearing H, methyl, vinyl, ethynyl or phenyl groups) were prepared by PCR using the corresponding base-modified 2'-deoxyribonucleoside triphosphates (dNTPs). The modified templates were used in an in vitro transcription assay using RNA polymerase from Bacillus subtilis and Escherichia coli. Some modified nucleobases bearing smaller modifications (H, Me in 7-deazapurines) were perfectly tolerated by both enzymes, whereas bulky modifications (Ph at any nucleobase) and, surprisingly, uracil blocked transcription. Some middle-sized modifications (vinyl or ethynyl) were partly tolerated mostly by the E. coli enzyme. In all cases where the transcription proceeded, full length RNA product with correct sequence was obtained indicating that the modifications of the template are not mutagenic and the inhibition is probably at the stage of initiation. The results are promising for the development of bioorthogonal reactions for artificial chemical switching of the transcription.

Název v anglickém jazyce

Influence of major-groove chemical modifications of DNA on transcription by bacterial RNA polymerases

Popis výsledku anglicky

DNA templates containing a set of base modifications in the major groove (5-substituted pyrimidines or 7-substituted 7-deazapurines bearing H, methyl, vinyl, ethynyl or phenyl groups) were prepared by PCR using the corresponding base-modified 2'-deoxyribonucleoside triphosphates (dNTPs). The modified templates were used in an in vitro transcription assay using RNA polymerase from Bacillus subtilis and Escherichia coli. Some modified nucleobases bearing smaller modifications (H, Me in 7-deazapurines) were perfectly tolerated by both enzymes, whereas bulky modifications (Ph at any nucleobase) and, surprisingly, uracil blocked transcription. Some middle-sized modifications (vinyl or ethynyl) were partly tolerated mostly by the E. coli enzyme. In all cases where the transcription proceeded, full length RNA product with correct sequence was obtained indicating that the modifications of the template are not mutagenic and the inhibition is probably at the stage of initiation. The results are promising for the development of bioorthogonal reactions for artificial chemical switching of the transcription.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EE - Mikrobiologie, virologie

OECD FORD obor

—

Projekt

<a href="/cs/project/GA14-04289S" target="_blank" >GA14-04289S: Modifikace a bioorthogonální reakce ve velkém žlábku DNA pro regulaci vazby proteinů a genové exprese</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2016

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Nucleic Acids Research

ISSN

0305-1048

e-ISSN

—

Svazek periodika

44

Číslo periodika v rámci svazku

7

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

13

Strana od-do

3000-3012

Kód UT WoS článku

000375800200013

EID výsledku v databázi Scopus

2-s2.0-84965131464