Characterization of an ATP-dependent DNA ligase from the acidophilic archaeon "Ferroplasma acidarmanus" Fer1

Jackson, Brian R., Noble, Catherine, Lavesa-Curto, Manuel, Bond, Philip L. and Bowater, Richard P. (2007) Characterization of an ATP-dependent DNA ligase from the acidophilic archaeon "Ferroplasma acidarmanus" Fer1. Extremophiles, 11 2: 315-327. doi:10.1007/s00792-006-0041-2

Author Jackson, Brian R.
Noble, Catherine
Lavesa-Curto, Manuel
Bond, Philip L.
Bowater, Richard P.
Title Characterization of an ATP-dependent DNA ligase from the acidophilic archaeon "Ferroplasma acidarmanus" Fer1
Journal name Extremophiles   Check publisher's open access policy
ISSN 1431-0651
Publication date 2007
Sub-type Article (original research)
DOI 10.1007/s00792-006-0041-2
Volume 11
Issue 2
Start page 315
End page 327
Total pages 13
Place of publication Tokyo
Publisher Springer Japan
Language eng
Subject 0601 Biochemistry and Cell Biology
Formatted abstract
Analysis of the genome of “Ferroplasma acidarmanus” Fer1, an archaeon that is an extreme acidophile, identified an open reading frame encoding a putative ATP-dependent DNA ligase, which we termed FaLig. The deduced amino acid sequence of FaLig contains 595 amino acids, with a predicted molecular mass of 67.8 kDa. “F. acidarmanus” Fer1 is classified as a Euryarchaeote, but phylogenetic analysis using amino acid sequences showed that FaLig is more similar to DNA ligases from Crenarchaeota, suggesting that lateral transfer of these genes has occurred among archaea. The gene sequence encoding FaLig was cloned into a bacterial expression vector harbouring an upstream His-tag to aid purification. Conditions for expression and purification from Escherichia coli were identified and recombinant FaLig was confirmed to be an ATP-dependent DNA ligase. Optimal conditions for nick-joining by the protein were pH 6–7, 0.5 mM ATP, in the presence of either Mg2+ or Mn2+. Using a range of nicked, double-stranded nucleic acids, ligation was detected with the same substrates as previously determined for other DNA ligases. Although FaLig is the DNA ligase from one of the most extreme acidophilic organism yet studied, this characterization suggests that its biochemical mechanism is analogous to that of enzymes from other cellular systems.
Keyword Biochemistry & Molecular Biology
DNA ligase
Ferroplasma acidarmanus
DNA nick-joining
Bacteriophage-t4 Rna Ligase-2
Acid-mine Drainage
Genome Sequence
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Chemistry and Molecular Biosciences
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Created: Mon, 18 Feb 2008, 16:21:46 EST