Type IV-Like Pili Facilitate Transformation in Naturally Competent Archaea

Abstract

Naturally competent organisms are capable of DNA uptake directly from the environment through the process of transformation. Despite the importance of transformation to microbial evolution, DNA uptake remains poorly characterized outside of the bacterial domain. Here, we identify the pilus as a necessary component of the transformation machinery in archaea. We describe two naturally competent organisms, Methanococcus maripaludis and Methanoculleus thermophilus. In M. maripaludis, replicative vectors were transferred with an average efficiency of 2.4 × 103 transformants μg−1 DNA. In M. thermophilus, integrative vectors were transferred with an average efficiency of 2.7 × 103 transformants μg−1 DNA. Additionally, natural transformation of M. thermophilus could be used to introduce chromosomal mutations. To our knowledge, this is the first demonstration of a method to introduce targeted mutations in a member of the order Methanomicrobiales. For both organisms, mutants lacking structural components of the type IV-like pilus filament were defective for DNA uptake, demonstrating the importance of pili for natural transformation. Interestingly, competence could be induced in a noncompetent strain of M. maripaludis by expressing pilin genes from a replicative vector. These results expand the known natural competence pili to include examples from the archaeal domain and highlight the importance of pili for DNA uptake in diverse microbial organisms.

Document Details

Document Type
Pub Defense Publication
Publication Date
Aug 17, 2020
Source ID
10.1128/jb.00355-20

Entities

People

  • Dallas R. Fonseca
  • Kyle C Costa
  • Matthew P. Holten
  • Mohd Farid Abdul Halim

Organizations

  • Army Research Office
  • National Science Foundation
  • United States Department of Energy

Tags

Fields of Study

  • Biology
  • Environmental science

Readers

  • Agent-Based Social Robotics and Mobile-Assisted Learning in Virtual Environments.
  • Molecular Genetics

Technology Areas

  • Biotechnology
  • Biotechnology - Bioremediation