ArticleBMC molecular biology2006
Biochemical characterisation of LigN, an NAD+-dependent DNA ligase from the halophilic euryarchaeon Haloferax volcanii that displays maximal in vitro activity at high salt concentrations.
Article in BMC molecular biology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 36 citations in OpenAlex.
- Characterisation and engineering of a thermophilic RNA ligase from Palaeococcus pacificus.Nucleic acids research · 2024Article
- Microbial communities in the Dead Sea and their potential biotechnological applications.Communicative & integrative biology · 2024Review
- Codon usage bias analysis of the gene encoding NADArchives of microbiology · 2023Article
- Microbial adaptation to different environmental conditions: molecular perspective of evolved genetic and cellular systems.Archives of microbiology · 2022Review
- Experimental study of proteome halophilicity using nanoDSF: a proof of concept.Extremophiles : life under extreme conditions · 2021Article
- Review
- Structural adaptations of octaheme nitrite reductases from haloalkaliphilic Thioalkalivibrio bacteria to alkaline pH and high salinity.PloS one · 2017Article
- Biochemical and Structural Characterisation of DNA Ligases from Bacteria and Archaea.Bioscience reports · 2016Article
- Kinetic mechanism and fidelity of nick sealing by Escherichia coli NAD+-dependent DNA ligase (LigA).Nucleic acids research · 2016Article
- Archaeal Nucleic Acid Ligases and Their Potential in Biotechnology.Archaea (Vancouver, B.C.) · 2015Review
- The haloarchaeal MCM proteins: bioinformatic analysis and targeted mutagenesis of the β7-β8 and β9-β10 hairpin loops and conserved zinc binding domain cysteines.Frontiers in microbiology · 2014Article
- Broad nucleotide cofactor specificity of DNA ligase from the hyperthermophilic crenarchaeon Hyperthermus butylicus and its evolutionary significance.Extremophiles : life under extreme conditions · 2013Article
- Function and biotechnology of extremophilic enzymes in low water activity.Aquatic biosystems · 2012Article
- DNA binding in high salt: analysing the salt dependence of replication protein A3 from the halophile Haloferax volcanii.Archaea (Vancouver, B.C.) · 2012Article
- Uracil-DNA glycosylase of Thermoplasma acidophilum directs long-patch base excision repair, which is promoted by deoxynucleoside triphosphates and ATP/ADP, into short-patch repair.Journal of bacteriology · 2011Article
- Halophilic enzyme activation induced by salts.Scientific reports · 2011Article
- Structural basis for the aminoacid composition of proteins from halophilic archea.PLoS biology · 2009Article
- A single amino acid substitution in the DNA-binding domain of Aeropyrum pernix DNA ligase impairs its interaction with proliferating cell nuclear antigen.Extremophiles : life under extreme conditions · 2007Article
- Structure and function of cold shock proteins in archaea.Journal of bacteriology · 2007Article
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2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundDNA ligases are required for DNA strand joining in all forms of cellular life. NAD+-dependent DNA ligases are found primarily in eubacteria but also in some eukaryotic viruses, bacteriophage and archaea. Among the archaeal NAD+-dependent DNA ligases is the LigN enzyme of the halophilic euryarchaeon Haloferax volcanii, the gene for which was apparently acquired by Hfx. volcanii through lateral gene transfer (LGT) from a halophilic eubacterium. Genetic studies show that the LGT-acquired LigN enzyme shares an essential function with the native Hfx. volcanii ATP-dependent DNA ligase protein LigA.
resultsTo characterise the enzymatic properties of the LigN protein, wild-type and three mutant forms of the LigN protein were separately expressed in recombinant form in E.coli and purified to apparent homogeneity by immobilised metal ion affinity chromatography (IMAC). Non-isotopic DNA ligase activity assays using lambda DNA restriction fragments with 12 bp cos cohesive ends were used to show that LigN activity was dependent on addition of divalent cations and salt. No activity was detected in the absence of KCl, whereas maximum activity could be detected at 3.2 M KCl, close to the intracellular KCl concentration of Hfx. volcanii cells.
conclusionLigN is unique amongst characterised DNA ligase enzymes in displaying maximal DNA strand joining activity at high (> 3 M) salt levels. As such the LigN enzyme has potential both as a novel tool for biotechnology and as a model enzyme for studying the adaptation of proteins to high intracellular salt levels.
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