CPL - Chalmers Publication Library
| Utbildning | Forskning | Styrkeområden | Om Chalmers | In English In English Ej inloggad.

Discrimination against the Cytosine Analog tC by Escherichia coli DNA Polymerase IV DinB

J. M. Walsh ; I. Bouamaied ; T. Brown ; Marcus Wilhelmsson (Institutionen för kemi- och bioteknik, Fysikalisk kemi) ; P. J. Beuning
Journal of Molecular Biology (0022-2836). Vol. 409 (2011), 2, p. 89-100.
[Artikel, refereegranskad vetenskaplig]

The cytosine analog 1,3-diaza-2-oxophenothiazine (tC) is a fluorescent nucleotide that forms Watson Crick base pairs with dG. The Klenow fragment of DNA polymerase I (an A-family polymerase) can efficiently bypass tC on the template strand and incorporate deoxyribose-triphosphate-tC into the growing primer terminus. Y-family DNA polymerases are known for their ability to accommodate bulky lesions and modified bases and to replicate beyond such nonstandard DNA structures in a process known as translesion synthesis. We probed the ability of the Escherichia coli Y-family DNA polymerase DinB (Pol IV) to copy DNA containing tC and to incorporate tC into a growing DNA strand. DinB selectively adds dGTP across from tC in template DNA but cannot extend beyond the newly formed G:tC base pair. However, we find that DinB incorporates the tC deoxyribonucleotide triphosphate opposite template G and extends from tC. Therefore, DinB displays asymmetry in terms of its ability to discriminate against the modification of the DNA template compared to the incoming nucleotide. In addition, our finding that DinB (a lesion-bypass DNA polymerase) specifically discriminates against tC in the template strand may suggest that DinB discriminates against template modifications in the major groove of DNA.

Nyckelord: primer extension, tryptophan fluorescence binding assay, major groove, translesion synthesis



Den här publikationen ingår i följande styrkeområden:

Läs mer om Chalmers styrkeområden  

Denna post skapades 2011-06-27. Senast ändrad 2015-08-25.
CPL Pubid: 142515

 

Läs direkt!


Länk till annan sajt (kan kräva inloggning)


Institutioner (Chalmers)

Institutionen för kemi- och bioteknik, Fysikalisk kemi (2005-2014)

Ämnesområden

Livsvetenskaper
Nanovetenskap och nanoteknik
Fysikalisk kemi
Biofysikalisk kemi
Kinetik
Spektroskopi
Molekylärbiologi
Molekylär biofysik

Chalmers infrastruktur