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An Extended Network of Genomic Maintenance in the Archaeon Pyrococcus abyssi Highlights Unexpected Associations between Eucaryotic Homologs ArchiMer
Pluchon, Pierre Francois; Fouqueau, Thomas; Creze, Christophe; Laurent, Sebastien; Briffotaux, Julien; Hogrel, Gaelle; Palud, Adeline; Henneke, Ghislaine; Godfroy, Anne; Hausner, Winfried; Thomm, Michael; Nicolas, Jacques; Flament, Didier.
In Archaea, the proteins involved in the genetic information processing pathways, including DNA replication, transcription, and translation, share strong similarities with those of eukaryotes. Characterizations of components of the eukaryotic-type replication machinery complex provided many interesting insights into DNA replication in both domains. In contrast, DNA repair processes of hyperthermophilic archaea are less well understood and very little is known about the intertwining between DNA synthesis, repair and recombination pathways. The development of genetic system in hyperthermophilic archaea is still at a modest stage hampering the use of complementary approaches of reverse genetics and biochemistry to elucidate the function of new candidate DNA...
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Ano: 2013 URL: http://archimer.ifremer.fr/doc/00171/28198/26556.pdf
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Caractérisation des ADN polymérases de Pyrococcus abyssi en présence de matrices ADN endommagées ArchiMer
Palud, Adeline.
Hyperthermophilic archaea cope with harsh conditions such as high temperatures, high pressures, pH shifts and ionising radiations. Such environments favour the apparition of DNA lesions like apurinic (AP) sites or oxidized DNA bases (8-oxo-dG and 8-oxo-dA). Here, Pyrococcus abyssi (Pab) was used as an attractive model to analyse the impact of such lesions onto the maintenance of genome integrity. The first part aimed at detecting DNA damages in the genome of Pab: -Endogenous AP sites and the oxidized DNA base 8-oxo-dG persist at a slightly higher level in Pab genome compared with Escherichia coli. -Under oxydative stress conditions performed in a gas-lift bioreactor for the first time, the 8-oxo-dG increasing rate is correlated to the important cell...
Tipo: Text Palavras-chave: PCR on damaged DNA; Gas lift bioreactor; Detection of lesions; Translesion; 8 oxo dGTP; 8 oxo dA; 8 oxo dG; AP sites; Archaea; DNA polymerases; PCR sur ADN endommagé; Bioréacteur gas lift; Détection de lésions; Translésion; 8 oxo dGTP; 8 oxo dA; 8 oxo dG; Sites AP; Archaea; ADN polymérases.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/these-6931.pdf
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Intrinsic properties of the two replicative DNA polymerases of Pyrococcus abyssi in replicating abasic sites: possible role in DNA damage tolerance? ArchiMer
Palud, Adeline; Villani, Giuseppe; L'Haridon, Stephane; Querellou, Joel; Raffin, Jean-paul; Henneke, Ghislaine.
Spontaneous and induced abasic sites in hyperthermophiles DNA have long been suspected to occur at high frequency. Here, Pyrococcus abyssi was used as an attractive model to analyse the impact of such lesions onto the maintenance of genome integrity. We demonstrated that endogenous AP sites persist at a slightly higher level in P. abyssi genome compared with Escherichia coli. Then, the two replicative DNA polymerases, PabpolB and PabpolD, were characterized in presence of DNA containing abasic sites. Both Pabpols had abortive DNA synthesis upon encountering AP sites. Under running start conditions, PabpolB could incorporate in front of the damage and even replicate to the full-length oligonucleotides containing a specific AP site, but only when present at...
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Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/publication-6113.pdf
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Structure of an octameric form of the minichromosome maintenance protein from the archaeon Pyrococcus abyssi ArchiMer
Cannone, Giuseppe; Visentin, Silvia; Palud, Adeline; Henneke, Ghislaine; Spagnolo, Laura.
Cell division is a complex process that requires precise duplication of genetic material. Duplication is concerted by replisomes. The Minichromosome Maintenance (MCM) replicative helicase is a crucial component of replisomes. Eukaryotic and archaeal MCM proteins are highly conserved. In fact, archaeal MCMs are powerful tools for elucidating essential features of MCM function. However, while eukaryotic MCM2-7 is a heterocomplex made of different polypeptide chains, the MCM complexes of many Archaea form homohexamers from a single gene product. Moreover, some archaeal MCMs are polymorphic, and both hexameric and heptameric architectures have been reported for the same polypeptide. Here, we present the structure of the archaeal MCM helicase from Pyrococcus...
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Ano: 2017 URL: http://archimer.ifremer.fr/doc/00373/48434/48701.pdf
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The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea ArchiMer
Killelea, Tom; Palud, Adeline; Akcha, Farida; Lemor, Mélanie; L'Haridon, Stephane; Godfroy, Anne; Henneke, Ghislaine.
8-oxodeoxyguanosine (8-oxodG), a major oxidised base modification, has been investigated to study its impact on DNA replication in hyperthermophilic Archaea. Here we show that 8-oxodG is formed in the genome of growing cells, with elevated levels following exposure to oxidative stress. Functional characterisation of cell-free extracts and the DNA polymerisation enzymes, PolB, PolD, and the p41/p46 complex, alone or in the presence of accessory factors (PCNA and RPA) indicates that translesion synthesis occurs under replicative conditions. One of the major polymerisation effects was stalling, but each of the individual proteins could insert and extend past 8-oxodG with differing efficiencies. The introduction of RPA and PCNA influenced PolB and PolD in...
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Ano: 2019 URL: https://archimer.ifremer.fr/doc/00502/61372/64989.pdf
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