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Diversity and three-dimensional structures of the alpha Mcr of the methanogenic Archaea from the anoxic region of Tucuruí Lake, in Eastern Brazilian Amazonia Genet. Mol. Biol.
Santana,Priscila Bessa; Ghilardi Junior,Rubens; Alves,Claudio Nahum; Silva,Jeronimo Lameira; McCulloch,John Anthony; Schneider,Maria Paula Cruz; Silva,Artur da Costa da.
Methanogenic archaeans are organisms of considerable ecological and biotechnological interest that produce methane through a restricted metabolic pathway, which culminates in the reaction catalyzed by the Methyl-coenzyme M reductase (Mcr) enzyme, and results in the release of methane. Using a metagenomic approach, the gene of the a subunit of mcr (mcrα) was isolated from sediment sample from an anoxic zone, rich in decomposing organic material, obtained from the Tucuruí hydroelectric dam reservoir in eastern Brazilian Amazonia. The partial nucleotide sequences obtained were 83 to 95% similar to those available in databases, indicating a low diversity of archaeans in the reservoir. Two orders were identified -the Methanomicrobiales, and a unique Operational...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Amazonian; Archaea; Diversity; Methyl-coenzyme M reductase; Mcrα; Homology modeling.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000100018
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La réplication de l'ADN chez l'euryarchaea Pyrococcus Abyssi : mise en place et dynamique du complexe ArchiMer
Rouillon, Christophe.
DNA replication occurs through a large protein complex assembly called replisome. An understanding of its structure and function requires its in vitro reassembly from individual subunits. In all living organisms, the elongation of a primed DNA template by DNA polymerase, with high processivity, requires the accessory protein DNA sliding clamp (PCNA) that is loaded onto DNA by the clamp loader (RF-C). Generally, the archaeal proteins involved in DNA replication are more similar to those of the higher organisms (Eukarya) rather than to those from Bacteria. This study have focussed on the PCNA loading mechanism and on the DNA synthesis in the hyperthermophilic Euryarchaea Pyrococcus abyssi. To investigate the importance of some protein-protein and...
Tipo: Text Palavras-chave: Pyrococcus; Archaea; DNA synthesis; Interaction; RP A; RF C; PCNA; DNA polymerase; DNA replication; Pyrococcus; Archée; Archaea; Synthese d'ADN; Interaction; RP A; RF C; PCNA; ADN polymerase; Replication de l'ADN.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/these-2461.pdf
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Role of RadA and DNA Polymerases in Recombination-Associated DNA Synthesis in Hyperthermophilic Archaea ArchiMer
Hogrel, Gaelle; Lu, Yang; Alexandre, Nicolas; Bossé, Audrey; Dulermo, Remi; Ishino, Sonoko; Ishino, Yoshizumi; Flament, Didier.
Among the three domains of life, the process of homologous recombination (HR) plays a central role in the repair of double-strand DNA breaks and the restart of stalled replication forks. Curiously, main protein actors involved in the HR process appear to be essential for hyperthermophilic Archaea raising interesting questions about the role of HR in replication and repair strategies of those Archaea living in extreme conditions. One key actor of this process is the recombinase RadA, which allows the homologous strand search and provides a DNA substrate required for following DNA synthesis and restoring genetic information. DNA polymerase operation after the strand exchange step is unclear in Archaea. Working with Pyrococcus abyssi proteins, here we show...
Tipo: Text Palavras-chave: Homologous recombination; Archaea; DNA polymerase; Recombinase; DNA repair.
Ano: 2020 URL: https://archimer.ifremer.fr/doc/00641/75274/75539.pdf
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The many ways of coping with pressure ArchiMer
Oger, Philippe M.; Jebbar, Mohamed.
The current paper reviews strategies employed by microorganisms from the deep biosphere, especially piezophiles (from the greek piezo = to press and philo = love), to cope with high hydrostatic pressure (HHP) prevailing in these biotopes. The aim of this review is not to constitute an exhaustive report of our current knowledge on the physiology of piezophiles, as recent reviews have covered part of this subject in detail (Abe, 2007; Lauro and Bartlett, 2008; Michiels et al., 2008; Simonato et al., 2006). Rather, we illustrate here, via a few chosen examples, where we stand in our understanding of the mechanisms employed by microorganisms from the depths of our planet to cope with HHP. (C) 2010 Institut Pasteur. Published by Elsevier Masson SAS. All rights...
Tipo: Text Palavras-chave: Adaptation and evolution; Archaea; Deep biosphere; High hydrostatic pressure; Piezophile; Stress response.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00027/13797/11043.pdf
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DNA polymerase switching on homotrimeric PCNA at the replication fork of the euryarchaea Pyrococcus abyssi ArchiMer
Rouillon, Christophe; Henneke, Ghislaine; Flament, Didier; Querellou, Joel; Raffin, Jean-paul.
DNA replication in Archaea, as in other organisms, involves large protein complexes called replisomes. In the Euryarchaeota subdomain, only two putative replicases have been identified, and their roles in leading and lagging strand DNA synthesis are still poorly understood. In this study, we focused on the coupling of proliferating cell nuclear antigen (PCNA)loading mechanisms with DNA polymerase function in the Euryarchaea Pyrococcus abyssi. PCNA spontaneously loaded onto primed DNA, and replication factor C dramatically increased this loading. Surprisingly, the family B DNA polymerase (Pol B) also increased PCNA loading, probably by stabilizing the clamp on primed DNA via an essential motif. In contrast, on an RNA-primed DNA template, the PCNA/Pol B...
Tipo: Text Palavras-chave: RF C; PCNA loading; DNA polymerase switching; DNA replication; Archaea.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-2638.pdf
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Les ADN polymérases B et D de l'Archaea hyperthermophile, Pyrococcus abyssi : contribution à l'étude des relations structure-fonction ArchiMer
Castrec, Benoit.
DNA replication is a functionally conserved mechanism among the three domains of life. This processus is performed by DNA polymerases with two accessories factors, PCNA and RF-C, sliding clamp and clamp loader, respectively. Our model of study, the Euryarchaeota Pyrococcus abyssi, possesses one monomeric DNA polymerase, belonging to the Family B, and one heterodimeric DNA polymerase, belonging to the Family D. Usually, proteins interact with PCNA at a hydrophobic pocket which is formed by the IDCL (interdomain connecting loop) and the C-terminus. These interactions are mediated through common motifs like the PIP box (motif PCNA interacting Protein). We have demonstrated that Pol B has just one PIP box motif at the C-terminus, while Pol D has two PIP-type...
Tipo: Text Palavras-chave: Thermostability; Interactions; Motifs; DNA replication; RF C; PCNA; DNA polymerases; Archaea; Thermostabilité; Interactions; Motifs; Réplication de l'ADN; RF C; PCNA; ADN polymérases; Archaea.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/these-6940.pdf
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Binding to PCNA in Euryarchaeal DNA Replication Requires Two PIP Motifs for DNA Polymerase D and One PIP Motif for DNA Polymerase B ArchiMer
Castrec, Benoit; Rouillon, Christophe; Henneke, Ghislaine; Flament, Didier; Querellou, Joel; Raffin, Jean-paul.
Replicative DNA polymerases possess a canonical C-terminal proliferating cell nuclear antigen (PCNA)-binding motif termed the PCNA-interacting protein (PIP) box. We investigated the role of the PIP box on the functional interactions of the two DNA polymerases, PabPol B (family B) and PabPol D (family D), from the hyperthermophilic euryarchaeon Pyrococcus abyssi, with its cognate PCNA. The PIP box was essential for interactions of PabPol B with PCNA, as shown by surface plasmon resonance and primer extension studies. In contrast, binding of PabPol D to PCNA was affected only partially by removing the PIP motif. We identified a second palindromic PIP box motif at the N-terminus of the large subunit of PabPol D that was required for the interactions of PabPol...
Tipo: Text Palavras-chave: Archaea; PIP box; PCNA binding motifs; DNA polymerases; DNA replication.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-7317.pdf
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PCR performance of a thermostable heterodimeric archaeal DNA polymerase ArchiMer
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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Subseafloor archaeal communities: from the surface to a deep hot biosphere? ArchiMer
Roussel, Erwan.
The sub-seafloor biosphere may contain two thirds of Earth's total prokaryotic biomass. The large and active microbial populations buried in the sub-seafloor play a key role in global biogeochemical cycles. However, little is known about these prokaryotic communities. The depth limit of this sub-seafloor biosphere is still unreached, and elevated temperatures as well as insufficient energy sources are the likely factors limiting life at depth. Interestingly, archaeal communities with unknown physiologies and no cultured relatives seem to have a key role in deep marine sediments and hydrothermal ecosystems. As most deep biosphere microorganisms detected so far have been extremely resistant to cultivation, molecular approaches remain to date the most...
Tipo: Text Palavras-chave: Sediment; Hydrothermal system; Ultramafic; CM DGGE; DGGE; PmoA; DsrA; McrA; AmoA; 16SrRNA; Bacteria; Archaea; Deep biosphere; Sédiment; Écosystème hydrothermal; Ultramafique; CM DGGE; DGGE; PmoA; DsrA; McrA; AmoA; 16SrRNA; Bacteria; Archaea; Biosphère profonde.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/these-4630.pdf
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Maintenance génomique chez l'Archaea hyperthermophile Pyrococcus abyssi : découverte de nouvelles interactions physiques et caractérisation fonctionnelle ArchiMer
Briffotaux, Julien.
Archaea are micro-organisms present in all ecosystems, but are majoritary present in extremophilic environments. Hyperthermophilic Archaea, as Pyrococcus abyssi, are permanently exposed at temperature which can increase the rate of spontaneous DNA damage. It is probable that hyperthermophilic Archaea possess efficient molecular mechanism to duplicate, maintain and stabilize their genomes. The aim of this project was to investigate the interaction network involved in the process of DNA replication and DNA repair. The methodological approach consists in the coupling of pull-down with mass spectrometry identification of interacting partners. A preliminary interaction network was identified that was composed of new proteins as well as unsuspected interactions...
Tipo: Text Palavras-chave: Protein Protein Interaction; DNA Replication; Genomic Maintenance; Archaea; Interaction Protéine Protéine; Réplication de l'ADN; Maintenance Génomique; Archaea.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/these-3722.pdf
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Differential Activities of DNA Polymerases in Processing Ribonucleotides during DNA Synthesis in Archaea ArchiMer
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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The PCNA from Thermococcus fumicolans functionally interacts with DNA polymerase delta ArchiMer
Henneke, Ghislaine; Raffin, Jean-paul; Ferrari, Elena; Jonsson, Zophonías; Dietrich, Jacques; Hubscher, Ulrich.
We have cloned the gene encoding proliferating cell nuclear antigen (PCNA) from the hyperthermophilic euryarchaeote Thermococcus fumicolans (Tfu). Tfu PCNA contains 250 amino acids with a calculated M-r of 28,000 and is 26% identical to human PCNA. Next, Tfu PCNA was overexpressed in Escherichia coli and it showed an apparent molecular mass of 33.5 kDa. The purified Tfu PCNA was tested first with recombinant Tfu DNA polymerase I (Tfu pol) and second with calf thymus DNA polymerase delta (pol delta). When tested with the homologous Tfu pol on bacteriophage lambda DNA, large amounts of Tfu PCNA were required to obtain two- to threefold stimulation. Surprisingly, however, Tfu PCNA was much more efficient than human PCNA in stimulating calf thymus pol delta....
Tipo: Text Palavras-chave: DNA polymerase delta; Conservation; Thermococcus fumicolans; Archaea; Hyperthermophile; PCNA.
Ano: 2000 URL: http://archimer.ifremer.fr/doc/2000/publication-659.pdf
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Replication slippage of the thermophilic DNA polymerases B and D from the Euryarchaeota Pyrococcus abyssi ArchiMer
Castillo-lizardo, Melissa; Henneke, Ghislaine; Viguera, Enrique.
Replication slippage or slipped-strand mispairing involves the misalignment of DNA strands during the replication of repeated DNA sequences, and can lead to genetic rearrangements such as microsatellite instability. Here, we show that PolB and PolD replicative DNA polymerases from the archaeal model Pyrococcus abyssi (Pab) slip in vitro during replication of a single-stranded DNA template carrying a hairpin structure and short direct repeats. We find that this occurs in both their wild-type (exo+) and exonuclease deficient (exo-) forms. The slippage behavior of PabPolB and PabPolD, probably due to limited strand displacement activity, resembles that observed for the high fidelity P furiosus (Pfu) DNA polymerase. The presence of PabPCNA inhibited PabPolB...
Tipo: Text Palavras-chave: Slippage; Primer-template misalignment; DNA polymerases; Strand displacement activity; Archaea.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00212/32375/30835.pdf
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Archaeal Methane Cycling Communities Associated with Gassy Subsurface Sediments of Marennes-Oleron Bay (France) ArchiMer
Roussel, Erwan; Sauvadet, Anne Laure; Allard, Jonathan; Chaduteau, Carine; Richard, Pierre; Cambon Bonavita, Marie-anne; Chaumillon, Eric.
In Marennes-Oleron Bay, a macro-tidal bay located on the French Atlantic coast, kilometer-scale acoustic turbidity reveals an accumulation of free gas in the sediment. Large concentrations of organic matter and rapid sedimentation rates provide ideal settings for biogenic methane cycling. We integrate seismic, sedimentologic, biogeochemical and molecular genetic approaches to determine whether microbial methane cycling is involved in this process. Here we show that the acoustic turbidity upper boundary matched with X-ray facies displaying fissures with the highest methane concentrations, demonstrating the existence of methane bubbles in the sediment. 16S rRNA and mcrA gene clone libraries were dominated by sequences affiliated to the three known ANME...
Tipo: Text Palavras-chave: Sediment; Methane; McrA; 16S rRNA; Archaea.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6165.pdf
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Archaea in organic-lean and organic-rich marine subsurface sediments: an environmental gradient reflected in distinct phylogenetic lineages ArchiMer
Durbin, Alan M.; Teske, Andreas.
Examining the patterns of archaeal diversity in little-explored organic-lean marine subsurface sediments presents an opportunity to study the association of phylogenetic affiliation and habitat preference in uncultured marine Archaea. Here we have compiled and re-analyzed published archaeal 16S rRNA clone library datasets across a spectrum of sediment trophic states characterized by a wide range of terminal electron-accepting processes. Our results show that organic-lean marine sediments in deep marine basins and oligotrophic open ocean locations are inhabited by distinct lineages of archaea that are not found in the more frequently studied, organic-rich continental margin sediments. We hypothesize that different combinations of electron donor and acceptor...
Tipo: Text Palavras-chave: Archaea; Marine sediments; Oligotrophy; Subsurface; Phylogeny; Uncultured archaea.
Ano: 2012 URL: https://archimer.ifremer.fr/doc/00263/37444/35695.pdf
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Phased oscillations in cell numbers and nitrate in batch cultures of Alexandrium tamarense (Dinophyceae) ArchiMer
Collos, Yves; Hadjadji, Imene; Plisson, Benoit; Cecchi, Philippe; Laabir, Mohamed; Bechemin, Christian; Masseret, Estelle.
Alexandrium tamarense (M. Lebour) Balech strains isolated in spring 2007 from a single bloom in Thau lagoon have been grown in nonaxenic artificial media. For three strains showing large oscillations in biomass (crashes followed by recoveries) on a scale of several days, a significant relationship was observed between changes in cell densities (as in vivo fluorescence) and changes in nitrate concentrations. Increases in cell densities were accompanied by decreases in nitrate, while decreases in cell densities corresponded to increases in nitrate, presumably due to nitrification. Net increases in nitrate could reach up to 15 mu mol N . L(-1) . d(-1) indicating a very active nitrifying archaeal/bacterial population. However, following population crashes,...
Tipo: Text Palavras-chave: Alexandrium tamarense; Archaea; Bacteria; Decay; Growth; Nitrate; Nitrification; Oscillations.
Ano: 2011 URL: http://archimer.ifremer.fr/doc/00049/16046/13585.pdf
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Methanogenic activity and diversity in the centre of the Amsterdam Mud Volcano, Eastern Mediterranean Sea ArchiMer
Lazar, Cassandre; Parkes, R. John; Cragg, Barry A.; L'Haridon, Stephane; Toffin, Laurent.
Marine mud volcanoes are geological structures emitting large amounts of methane from their active centres. The Amsterdam mud volcano (AMV), located in the Anaximander Mountains south of Turkey, is characterized by intense active methane seepage produced in part by methanogens. To date, information about the diversity or the metabolic pathways used by the methanogens in active centres of marine mud volcanoes is limited. 14C-radiotracer measurements showed that methylamines/methanol, H2/CO2 and acetate were used for methanogenesis in the AMV. Methylotrophic methanogenesis was measured all along the sediment core, Methanosarcinales affiliated sequences were detected using archaeal 16S PCR-DGGE and mcrA gene libraries, and enrichments of methanogens showed...
Tipo: Text Palavras-chave: Methanogenesis; McrA; Archaea.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00087/19795/17538.pdf
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Comparison of microbial communities associated with three Atlantic ultramafic hydrothermal systems ArchiMer
Roussel, Erwan; Konn, Cecile; Charlou, Jean-luc; Donval, Jean-pierre; Fouquet, Yves; Querellou, Joel; Prieur, Daniel; Cambon-bonavita, Marie-anne.
The distribution of Archaea and methanogenic, methanotrophic and sulfate-reducing communities in three Atlantic ultramafic-hosted hydrothermal systems (Rainbow, Ashadze, Lost City) was compared using 16S rRNA gene and functional gene (mcrA, pmoA and dsrA) clone libraries. The overall archaeal community was diverse and heterogeneously distributed between the hydrothermal sites and the types of samples analyzed (seawater, hydrothermal fluid, chimney and sediment). The Lost City hydrothermal field, characterized by high alkaline warm fluids (pH>11; T<95 °C), harbored a singular archaeal diversity mostly composed of unaffiliated Methanosarcinales. The archaeal communities associated with the recently discovered Ashadze 1 site, one of the deepest active...
Tipo: Text Palavras-chave: Archaea; Hydrothermal vent; Mid-Atlantic Ridge; 16S rRNA gene; Sediment; Ultramafic.
Ano: 2011 URL: http://archimer.ifremer.fr/doc/00067/17836/15573.pdf
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A di-O-dihydrogeranylgeranyl glycerol from Thermococus S 557, a novel ether lipid, and likely intermediate in the biosynthesis of diethers in Archaea ArchiMer
Gonthier, Isabelle; Rager, Marie-noëlle; Metzger, Pierre; Guezennec, Jean; Largeau, Claude.
The lipids of a deep-sea hydrothermal vent arch ae on, Thermococcus S 557, were isolated, purified and structurally determined. Based on acid methanolysis and spectroscopic studies, the polar lipids were shown to comprise diphytanyl glycerol and dibiphytanyl diglycerol, typical membrane lipids of Arch aea. From the neutral lipids, 2,3-di-O-dihydro-14,15-geranylgeranyl glycerol was isolated. This novel ether lipid is, very likely, a close intermediate in the biosynthesis of diphytanyl glycerol diether in Arch aea.
Tipo: Text Palavras-chave: Mass spectrometry; NMR; Novel glycerol diether lipid; Thermococcus sp; Archaea.
Ano: 2001 URL: http://archimer.ifremer.fr/doc/2001/publication-736.pdf
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The heterodimeric primase from the euryarchaeon Pyrococcus abyssi: A multifunctional enzyme for initiation and repair? ArchiMer
Le Breton, Magali; Henneke, Ghislaine; Norais, C; Flament, Didier; Myllykallio, H; Querellou, Joel; Raffin, Jean-paul.
We report on the characterization of the DNA primase complex of the hyperthermophilic archaeon Pyrococcus abyssi (Pab). The Pab DNA primase complex is composed of the proteins Pabp41 and Pabp46, which show sequence similarities to the p49 and p58 subunits, respectively, of the eukaryotic polymerase alpha-primase complex. Both subunits were expressed, purified, and characterized. The Pabp41 subunit alone had no RNA synthesis activity but could synthesize long (up to 3 kb) DNA strands. Addition of the Pabp46 subunit increased the rate of DNA synthesis but decreased the length of the DNA fragments synthesized and conferred RNA synthesis capability. Moreover, in our experimental conditions, Pab DNA primase had comparable affinities for ribonucleotides and...
Tipo: Text Palavras-chave: Strand displacement; Gap filling; DNA primase; Archaea; DNA replication.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-3520.pdf
Registros recuperados: 38
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