Genomewide Insertional Mutagenesis in Streptomyces coelicolor Reveals Additional Genes Involved in Morphological Differentiation

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 97; no. 17; pp. 9642 - 9647
• Main Authors: Gehring, Amy M., Nodwell, Justin R., Beverley, Stephen M., Losick, Richard
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 15.08.2000; National Acad Sciences; National Academy of Sciences; The National Academy of Sciences
• Subjects: Amino Acid Sequence; Anti-Bacterial Agents - pharmacology; Bacteria; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - physiology; Base Sequence; Biological Sciences; Chromosomes; Chromosomes, Bacterial - genetics; DNA Transposable Elements - genetics; Genes; Genes, Bacterial - genetics; Genetic mutation; Genetic transposition; Genome, Bacterial; Hybridity; Microbial Sensitivity Tests; Microbiology; Molecular Sequence Data; Multigene Family - genetics; Mutagenesis, Insertional - genetics; Mutation; Nebramycin - analogs & derivatives; Nebramycin - pharmacology; Open reading frames; Phenotype; Phenotypes; Physical Chromosome Mapping; Pigments; Pigments, Biological - biosynthesis; Plasmids; Recombination, Genetic - genetics; Sequence Alignment; Spores, Bacterial - cytology; Spores, Bacterial - genetics; Spores, Bacterial - growth & development; Streptomyces - drug effects; Streptomyces - genetics; Streptomyces - growth & development; Streptomyces - metabolism; Streptomyces coelicolor; Transposons
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.170059797

The filamentous soil bacterium Streptomyces coelicolor undergoes a complex cycle of morphological differentiation involving the formation of an aerial mycelium and the production of pigmented antibiotics. We have developed a procedure for generating insertional mutants of S. coelicolor based on in vitro transposition of a plasmid library of cloned S. coelicolor DNAs. The insertionally mutated library was introduced into S. coelicolor, and transposon insertions were recovered at widely scattered locations around the chromosome. Many of the insertions revealed previously uncharacterized genes, and several caused novel mutant phenotypes, such as altered pigment production, enhanced antibiotic sensitivity, delayed or impaired formation of aerial hyphae, and a block in spore formation. The sporulation mutant harbored an insertion in one of three adjacent genes that are apparently unique to Streptomyces but are each represented by at least 20 paralogs at dispersed locations in the chromosome. Individual members of the three families often are found grouped together in a characteristic arrangement, suggesting that they have a common function.