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An extracellular, Ca 2+ -activated nuclease (EcnA) mediates transformation in a naturally competent archaeon.
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  • Kyle C. Costa,
  • Dallas R. Fonseca,
  • Leslie A. Day,
  • Kathryn K. Crone
Kyle C. Costa
University of Minnesota Twin Cities

Corresponding Author:kcosta@umn.edu

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Dallas R. Fonseca
University of Minnesota Twin Cities
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Leslie A. Day
University of Minnesota Twin Cities
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Kathryn K. Crone
University of Minnesota Twin Cities
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Abstract

Transformation, the uptake of DNA directly from the environment, is a major driver of gene flow in microbial populations. In bacteria, DNA uptake requires a nuclease that processes dsDNA to ssDNA, which is subsequently transferred into the cell and incorporated into the genome. However, the process of DNA uptake in archaea is still unknown. Previously, we cataloged genes essential to natural transformation in Methanococcus maripaludis, but few homologs of bacterial transformation-associated genes were identified. Here, we characterize one gene, MMJJ_16440 (named here as ecnA) to be a membrane-bound nuclease. We show that EcnA is Ca 2+-activated, present on the cell surface, and essential for transformation. While EcnA is capable of degrading several forms of DNA, the highest activity was observed with ssDNA as a substrate. Activity was also observed with circular dsDNA, suggesting that EcnA is an endonuclease. This is the first biochemical characterization of a transformation-associated protein in a member of the archaeal domain and suggests that both archaeal and bacterial transformation initiate in an analogous fashion.
Submitted to Molecular Microbiology
12 Feb 2024Review(s) Completed, Editorial Evaluation Pending
09 May 2024Assigned to Editor
09 May 2024Submission Checks Completed
11 Jun 2024Review(s) Completed, Editorial Evaluation Pending
11 Jun 2024Editorial Decision: Revise Minor
06 Aug 20242nd Revision Received
10 Aug 2024Submission Checks Completed
10 Aug 2024Assigned to Editor
11 Aug 2024Review(s) Completed, Editorial Evaluation Pending
11 Aug 2024Editorial Decision: Accept