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

0461200 - ÚOCHB 2017 RIV GB eng J - Journal Article
Raindlová, Veronika - Janoušková, Martina - Slavíčková, Michaela - Perlíková, Pavla - Boháčová, Soňa - Milisavljevič, Nemanja - Šanderová, Hana - Benda, Martin - Barvík, I. - Krásný, Libor - Hocek, Michal
Influence of major-groove chemical modifications of DNA on transcription by bacterial RNA polymerases.
Nucleic Acids Research. Roč. 44, č. 7 (2016), s. 3000-3012. ISSN 0305-1048. E-ISSN 1362-4962
R&D Projects: GA ČR GA14-04289S
Institutional support: RVO:61388963 ; RVO:61388971
Keywords : Escherichia coli RNA * Bacillus subtilis * restriction endonucleases
OECD category: Biochemistry and molecular biology; Microbiology (MBU-M)
Impact factor: 10.162, year: 2016
Method of publishing: Open access
http://nar.oxfordjournals.org/content/44/7/3000.full

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.
Permanent Link: http://hdl.handle.net/11104/0260868