Random transposon mutagenesis identifies genes essential for transformation in Methanococcus maripaludis

Random transposon mutagenesis identifies genes essential for transformation in Methanococcus maripaludis

Natural 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 underto...

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Bibliographic Details
Published in:Molecular genetics and genomics : MGG Vol. 298; no. 3; pp. 537 - 548
Main Authors: Fonseca, Dallas R., Loppnow, Madison B., Day, Leslie A., Kelsey, Elisa L., Abdul Halim, Mohd Farid, Costa, Kyle C.
Format: Journal Article
Language:English
Published: Berlin/Heidelberg Springer Berlin Heidelberg 01-05-2023
Springer Nature B.V
Subjects:
Animal Genetics and Genomics
Biochemistry
Biomedical and Life Sciences
Deoxyribonucleic acid
DNA
Gene flow
Genetic transformation
Genomes
Human Genetics
Life Sciences
Methanococcus - genetics
Methanococcus - metabolism
Methanococcus maripaludis
Microbial Genetics and Genomics
Mutagenesis
Mutagenesis - genetics
Mutagenesis, Insertional
Mutants
Original
Original Article
Plant Genetics and Genomics
Plasmids
Protein transport
Transposon mutagenesis
Transposons
Genetics
Competence
Transposon mutagenesis
Archaea
Natural transformation
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Summary:Natural 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.
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ISSN:1617-4615
1617-4623
DOI:10.1007/s00438-023-01994-7