ArticleProceedings of the National Academy of Sciences of the United States of America2020
Ranking essential bacterial processes by speed of mutant death.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 48 citations in OpenAlex.
- Applications of transposon-insertion sequencing for understanding bacterial physiology.Microbial genomics · 2026Review
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- Protein overabundance is driven by growth robustness.Science advances · 2026Article
- Comparative analysis of shared and unique mechanisms important for diverse strains of Pasteurella multocida to cause systemic infection in mice.PLoS pathogens · 2025Article
- Metabolic strategies that enable oral commensal persistence in a lower airway environment.mBio · 2025Article
- Protein overabundance is driven by growth robustness.bioRxiv : the preprint server for biology · 2025Article
- Inducible transposon mutagenesis identifies bacterial fitness determinants during infection in mice.Nature microbiology · 2025Article
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- Transposon mutagenesis screen ineLife · 2024Article
- Noise robustness and metabolic load determine the principles of central dogma regulation.Science advances · 2024Article
- Noise robustness and metabolic load determine the principles of central dogma regulation.ArXiv · 2024Article
- Noise robustness and metabolic load determine the principles of central dogma regulation.bioRxiv : the preprint server for biology · 2024Article
- Integrating bacterial molecular genetics with chemical biology for renewed antibacterial drug discovery.The Biochemical journal · 2024Review
- Maintenance of bacterial outer membrane lipid asymmetry: insight into MlaA.BMC microbiology · 2024Review
- Recent advances in genome-scale engineering inEngineering microbiology · 2024Review
- Article
- Genome-scale analysis of essential gene knockout mutants to identify an antibiotic target process.Antimicrobial agents and chemotherapy · 2023Article
- Microbial Primer: Transposon directed insertion site sequencing (TraDIS): A high throughput method for linking genotype to phenotype.Microbiology (Reading, England) · 2023Article
- Article
- Discovery of a glutathione utilization pathway in Francisella that shows functional divergence between environmental and pathogenic species.Cell host & microbe · 2023Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Mutant phenotype analysis of bacteria has been revolutionized by genome-scale screening procedures, but essential genes have been left out of such studies because mutants are missing from the libraries analyzed. Since essential genes control the most fundamental processes of bacterial life, this is a glaring deficiency. To address this limitation, we developed a procedure for transposon insertion mutant sequencing that includes essential genes. The method, called transformation transposon insertion mutant sequencing (TFNseq), employs saturation-level libraries of bacterial mutants generated by natural transformation with chromosomal DNA mutagenized heavily by in vitro transposition. The efficient mutagenesis makes it possible to detect large numbers of insertions in essential genes immediately after transformation and to follow their loss during subsequent growth. It was possible to order 45 essential processes based on how rapidly their inactivation inhibited growth. Inactivating ATP production, deoxyribonucleotide synthesis, or ribosome production blocked growth the fastest, whereas inactivating cell division or outer membrane protein synthesis blocked it the slowest. Individual mutants deleted of essential loci formed microcolonies of nongrowing cells whose sizes were generally consistent with the TFNseq ordering. The sensitivity of essential functions to genetic inactivation provides a metric for ranking their relative importance for bacterial replication and growth. Highly sensitive functions could represent attractive antibiotic targets since even partial inhibition should reduce growth.
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