Expanding the repertoire of DNA polymerase substrates: template-instructed incorporation of non-nucleoside triphosphate analogues by DNA polymerases beta and lambda.
We have recently shown that neither the base nor the sugar moieties of a nucleotide is an essential feature for its incorporation by DNA polymerases (pols) lambda and beta. Here we present the identification of novel non-nucleoside triphosphate (NNTP) derivatives belonging to three classes: (i) non-substrate-specific inhibitors of DNA pol lambda; (ii) substrate inhibitors which could preferentially be incorporated by either DNA pol lambda wild type or its Y505A mutant and (iii) the substrate inhibitor N-(Biphenylcarbonyl)-4-oxobutyl triphosphate which could be incorporated exclusively by DNA pol beta in a Mg2+-dependent manner, and preferentially pairs with A on the template. This compound represents the first example of a substrate lacking both nucleobase and ribose residue, showing distinct base-pairing properties with normal bases. Therefore, this NNTP analog can be considered as the prototype of an entirely novel class of DNA pol substrates.
Crespan
Emmanuele
E
Istituto di Genetica Molecolare, IGM-CNR, via Abbiategrasso 207, I-27100 Pavia, Italy.
Alexandrova
Ludmila
L
Khandazhinskaya
Anastasiya
A
Jasko
Maxim
M
Kukhanova
Marina
M
Villani
Giuseppe
G
Hübscher
Ulrich
U
Spadari
Silvio
S
Maga
Giovanni
G
D020029:Base Pairing; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D019951:DNA Polymerase beta / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry; D006801:Humans; D008274:Magnesium / Q000737:chemistry; D008345:Manganese / Q000737:chemistry; D009154:Mutation; D009705:Nucleosides / Q000737:chemistry; D010755:Organophosphates / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D013698:Templates, Genetic; D014443:Tyrosine / Q000235:genetics
D004791:Enzyme Inhibitors; C517045:N-(biphenylcarboxyl)-4-oxobutyl triphosphate; D009705:Nucleosides; D010755:Organophosphates; D014443:Tyrosine; D008345:Manganese; C109534:DNA polymerase X; C415367:DNA polymerase beta2; D004253:DNA Nucleotidylexotransferase; D019951:DNA Polymerase beta; D004259:DNA-Directed DNA Polymerase; D008274:Magnesium