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Provedor de dados:  ArchiMer
País:  France
Título:  Differential Activities of DNA Polymerases in Processing Ribonucleotides during DNA Synthesis in Archaea
Autores:  Lemor, Melanie
Kong, Ziqing
Henry, Etienne
Brizard, Raphael
Laurent, Sebastien
Bosse, Audrey
Henneke, Ghislaine
Data:  2018-12
Ano:  2018
Palavras-chave:  Archaea
DNA replication and repair
DNA polymerase
Nucleotide pool
Translesion synthesis
Resumo:  Consistent with the fact that ribonucleotides (rNTPs) are in excess over deoxyribonucleotides (dNTPs) in vivo, recent findings indicate that replicative DNA polymerases (DNA Pols) are able to insert ribonucleotides (rNMPs) during DNA synthesis, raising crucial questions about the fidelity of DNA replication in both Bacteria and Eukarya. Here, we report that the level of rNTPs is 20-fold higher than that of dNTPs in Pyrococcus abyssi cells. Using dNTP and rNTP concentrations present in vivo, we recorded rNMP incorporation in a template-specific manner during in vitro synthesis, with the family-D DNA Pol (PolD) having the highest propensity compared with the family-B DNA Pol and the p41/p46 complex. We also showed that ribonucleotides accumulate at a relatively high frequency in the genome of wild-type Thermococcales cells, and this frequency significantly increases upon deletion of RNase HII, the major enzyme responsible for the removal of RNA from DNA. Because ribonucleotides remain in genomic DNA, we then analyzed the effects on polymerization activities by the three DNA Pols. Depending on the identity of the base and the sequence context, all three DNA Pols bypass rNMP-containing DNA templates with variable efficiency and nucleotide (mis)incorporation ability. Unexpectedly, we found that PolD correctly base-paired a single ribonucleotide opposite rNMP-containing DNA templates. An evolutionary scenario is discussed concerning rNMP incorporation into DNA and genome stability.
Tipo:  Text
Idioma:  Inglês
Identificador:  https://archimer.ifremer.fr/doc/00464/57603/59796.pdf

https://archimer.ifremer.fr/doc/00464/57603/59797.pdf

DOI:10.1016/j.jmb.2018.10.004

https://archimer.ifremer.fr/doc/00464/57603/
Editor:  Academic Press Ltd- Elsevier Science Ltd
Formato:  application/pdf
Fonte:  Journal Of Molecular Biology (0022-2836) (Academic Press Ltd- Elsevier Science Ltd), 2018-12 , Vol. 430 , N. 24 , P. 4908-4924
Direitos:  info:eu-repo/semantics/openAccess

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