Abstract
AbstractNatural transformation, the process whereby a cell acquires DNA directly from the environment, is an important driver of evolution in microbial populations, yet the mechanism of DNA uptake is only characterized in bacteria. To expand our understanding of natural transformation in archaea, we undertook a genetic approach to identify a catalog of genes necessary for transformation in Methanococcus maripaludis. Using an optimized method to generate random transposon mutants, we screened 6144 mutant strains for defects in natural transformation and identified 25 transformation-associated candidate genes. Among these are genes encoding components of the type IV-like pilus, transcription/translation associated genes, genes encoding putative membrane bound transport proteins, and genes of unknown function. Interestingly, similar genes were identified regardless of whether replicating or integrating plasmids were provided as a substrate for transformation. Using allelic replacement mutagenesis, we confirmed that several genes identified in these screens are essential for transformation. Finally, we identified a homolog of a membrane bound substrate transporter in Methanoculleus thermophilus and verified its importance for transformation using allelic replacement mutagenesis, suggesting a conserved mechanism for DNA transfer in multiple archaea. These data represent an initial characterization of the genes important for transformation which will inform efforts to understand gene flow in natural populations. Additionally, knowledge of the genes necessary for natural transformation may assist in identifying signatures of transformation machinery in archaeal genomes and aid the establishment of new model genetic systems for studying archaea.
Funder
Division of Molecular and Cellular Biosciences
Publisher
Springer Science and Business Media LLC
Subject
Genetics,Molecular Biology,General Medicine
Reference62 articles.
1. Aas FE, Wolfgang M, Frye S et al (2002) Competence for natural transformation in Neisseria gonorrhoeae: components of DNA binding and uptake linked to type IV pilus expression. Mol Microbiol 46:749–760
2. Abdul Halim MF, Stoltzfus JD, Schulze S et al (2017) ArtA-dependent processing of a Tat substrate containing a conserved tripartite structure that is not localized at the C terminus. J Bacteriol 199:e00802-e816
3. Angelov A, Bergen P, Nadler F et al (2015) Novel Flp pilus biogenesis-dependent natural transformation. Front Microbiol 6:84
4. Ashton DM, Sweet GD, Somers JM, Kay WW (1980) Citrate transport in Salmonella typhimurium: studies with 2-fluoro-L-erythro-citrate as a substrate. Can J Biochem Physiol 58:797–803
5. Bao J, de Dios ME, Scheller S (2022) Efficient CRISPR/Cas12a-Based Genome-Editing Toolbox for Metabolic Engineering in Methanococcus maripaludis. ACS Synth Biol 11:2496–2503
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