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A Single Nucleotide Change in the polC DNA Polymerase III in Clostridium thermocellum Is Sufficient To Create a Hypermutator Phenotype

Lanahan, Anthony ; Zakowicz, Kamila ; Tian, Liang ; Olson, Daniel G ; Lynd, Lee R Buan, Nicole R ; Buan, Nicole R.

Applied and environmental microbiology, 2022-01, Vol.88 (1), p.e0153121-e0153121 [Periódico revisado por pares]

United States: American Society for Microbiology

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Título:
A Single Nucleotide Change in the polC DNA Polymerase III in Clostridium thermocellum Is Sufficient To Create a Hypermutator Phenotype
Autor: Lanahan, Anthony ; Zakowicz, Kamila ; Tian, Liang ; Olson, Daniel G ; Lynd, Lee R
Buan, Nicole R ; Buan, Nicole R.
Assuntos: Applied and Industrial Microbiology ; Base Composition ; BASIC BIOLOGICAL SCIENCES ; Biodiesel fuels ; Biofuels ; Cellulose ; Clostridium thermocellum ; Clostridium thermocellum - genetics ; CRISPR ; Deoxyribonucleic acid ; Directed evolution ; DNA ; DNA polymerase ; DNA Polymerase III ; DNA-directed DNA polymerase ; Editing ; Ethanol ; Evolution ; Evolution & development ; Evolutionary and Genomic Microbiology ; Genetic modification ; Metal ions ; Mutation ; Mutation rates ; Nucleotides ; Phenotype ; Phenotypes ; Phylogeny ; Proofreading ; RNA, Ribosomal, 16S ; Sequence Analysis, DNA ; Solubilization ; Transportation industry
É parte de: Applied and environmental microbiology, 2022-01, Vol.88 (1), p.e0153121-e0153121
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
AC02-05CH11231
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Citation Lanahan A, Zakowicz K, Tian L, Olson DG, Lynd LR. 2022. A single nucleotide change in the polC DNA polymerase III in Clostridium thermocellum is sufficient to create a hypermutator phenotype. Appl Environ Microbiol 88:e01531-21. https://doi.org/10.1128/AEM.01531-21.
Descrição: Clostridium thermocellum is a thermophilic, anaerobic bacterium that natively ferments cellulose to ethanol and is a candidate for cellulosic biofuel production. Recently, we identified a hypermutator strain of with a C669Y mutation in the gene, which encodes a DNA polymerase III enzyme. Here, we reintroduced this mutation using recently developed CRISPR tools to demonstrate that this mutation is sufficient to recreate the hypermutator phenotype. The resulting strain shows an approximately 30-fold increase in the mutation rate. This mutation is hypothesized to function by interfering with metal ion coordination in the PHP (polymerase and histidinol phosphatase) domain, which is responsible for proofreading. The ability to selectively increase the mutation rate in is a useful tool for future directed evolution experiments. Cellulosic biofuels are a promising approach to decarbonize the heavy-duty-transportation sector. A longstanding barrier to cost-effective cellulosic biofuel production is the recalcitrance of cellulose to solubilization. Native cellulose-consuming organisms, such as Clostridium thermocellum, are promising candidates for cellulosic biofuel production; however, they often need to be genetically modified to improve product formation. One approach is adaptive laboratory evolution. Our findings demonstrate a way to increase the mutation rate in this industrially relevant organism, which can reduce the time needed for adaptive evolution experiments.
Editor: United States: American Society for Microbiology
Idioma: Inglês

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A Single Nucleotide Change in the polC DNA Polymerase III in Clostridium thermocellum Is Sufficient To Create a Hypermutator Phenotype

Lanahan, Anthony ; Zakowicz, Kamila ; Tian, Liang ; Olson, Daniel G ; Lynd, Lee R Buan, Nicole R ; Buan, Nicole R.

United States: American Society for Microbiology

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