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| Lemor, Melanie; Kong, Ziqing; Henry, Etienne; Brizard, Raphael; Laurent, Sebastien; Bosse, Audrey; Henneke, Ghislaine. |
| 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... |
| Tipo: Text |
Palavras-chave: Archaea; DNA replication and repair; DNA polymerase; Nucleotide pool; Translesion synthesis. |
| Ano: 2018 |
URL: https://archimer.ifremer.fr/doc/00464/57603/59796.pdf |
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| Killelea, Tom; Ralec, Celine; Bosse, Audrey; Henneke, Ghislaine. |
| DNA polymerases are versatile tools used in numerous important molecular biological core technologies like the ubiquitous polymerase chain reaction (PCR), cDNA cloning, genome sequencing, and nucleic acid based diagnostics. Taking into account the multiple DNA amplification techniques in use, different DNA polymerases must be optimized for each type of application. One of the current tendencies is to reengineer or to discover new DNA polymerases with increased performance and broadened substrate spectra. At present, there is a great demand for such enzymes in applications, e.g., forensics or paleogenomics. Current major limitations hinge on the inability of conventional PCR enzymes, such as Taq, to amplify degraded or low amounts of template DNA. Besides,... |
| Tipo: Text |
Palavras-chave: DNA polymerase; Archaea; Family D; PCR; Pyrococcus. |
| Ano: 2014 |
URL: http://archimer.ifremer.fr/doc/00193/30450/28871.pdf |
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| Hogrel, Gaelle; Lu, Yang; Laurent, Sebastien; Henry, Etienne; Etienne, Clarisse; Duy Khanh Phung,; Dulermo, Remi; Bosse, Audrey; Pluchon, Pierre Francois; Clouet-d'Orval, Beatrice; Flament, Didier. |
| Several archaeal species prevalent in extreme environments are particularly exposed to factors likely to cause DNA damages. These include hyperthermophilic archaea (HA), living at temperatures >70°C, which arguably have efficient strategies and robust genome guardians to repair DNA damage threatening their genome integrity. In contrast to Eukarya and other archaea, homologous recombination appears to be a vital pathway in HA, and the Mre11–Rad50 complex exerts a broad influence on the initiation of this DNA damage response process. In a previous study, we identified a physical association between the Proliferating Cell Nuclear Antigen (PCNA) and the Mre11–Rad50 (MR) complex. Here, by performing co-immunoprecipitation and SPR analyses, we identified a... |
| Tipo: Text |
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| Ano: 2018 |
URL: http://archimer.ifremer.fr/doc/00439/55092/56534.pdf |
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