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Identification of novel factors involved in modulating motility of Salmonella enterica serotype typhimurium.

Bogomolnaya LM, Aldrich L, Ragoza Y, Talamantes M, Andrews KD, McClelland M, Andrews-Polymenis HL - PLoS ONE (2014)

Bottom Line: Mutants in 130 genes, not previously known to be involved in motility, had altered movement of at least one type, 9 mutants were severely impaired for both types of motility, while 33 mutants appeared defective on swimming motility plates but not swarming motility plates, and 49 mutants had reduced ability to move on swarming agar but not swimming agar.Both mutants that appeared non-motile and hypermotile on plates were assayed for expression levels of FliC and FljB on the bacterial surface and many of them had altered levels of these proteins.The phenotypes we report are the first phenotypes ever assigned to 74 of these open reading frames, as they are annotated as 'hypothetical genes' in the Typhimurium genome.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University, Bryan, Texas, United States of America; Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Russia.

ABSTRACT
Salmonella enterica serotype Typhimurium can move through liquid using swimming motility, and across a surface by swarming motility. We generated a library of targeted deletion mutants in Salmonella Typhimurium strain ATCC14028, primarily in genes specific to Salmonella, that we have previously described. In the work presented here, we screened each individual mutant from this library for the ability to move away from the site of inoculation on swimming and swarming motility agar. Mutants in genes previously described as important for motility, such as flgF, motA, cheY are do not move away from the site of inoculation on plates in our screens, validating our approach. Mutants in 130 genes, not previously known to be involved in motility, had altered movement of at least one type, 9 mutants were severely impaired for both types of motility, while 33 mutants appeared defective on swimming motility plates but not swarming motility plates, and 49 mutants had reduced ability to move on swarming agar but not swimming agar. Finally, 39 mutants were determined to be hypermotile in at least one of the types of motility tested. Both mutants that appeared non-motile and hypermotile on plates were assayed for expression levels of FliC and FljB on the bacterial surface and many of them had altered levels of these proteins. The phenotypes we report are the first phenotypes ever assigned to 74 of these open reading frames, as they are annotated as 'hypothetical genes' in the Typhimurium genome.

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Motility scores of single deletion mutants of Salmonella Typhimurium ATCC14028 on swimming (A) and swarming (B) agar.Stationary phase cultures were transferred to motility agar, and motility was scored after 3.5 and 5 hours of incubation at 37°C on swimming or swarming plates, respectively. Motility was scored on the scale from 0 to 10 with wild type motility equal to 5. Data are presented as average swimming or swarming score from experiments with triplicate samples, performed on three independent occasions. Dots located outside of shaded area indicate scores greater than 5.25 and lower than 1.25 to define mutants with increased or reduced (<25 % of wild type) motility.
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pone-0111513-g001: Motility scores of single deletion mutants of Salmonella Typhimurium ATCC14028 on swimming (A) and swarming (B) agar.Stationary phase cultures were transferred to motility agar, and motility was scored after 3.5 and 5 hours of incubation at 37°C on swimming or swarming plates, respectively. Motility was scored on the scale from 0 to 10 with wild type motility equal to 5. Data are presented as average swimming or swarming score from experiments with triplicate samples, performed on three independent occasions. Dots located outside of shaded area indicate scores greater than 5.25 and lower than 1.25 to define mutants with increased or reduced (<25 % of wild type) motility.

Mentions: We have screened a library of targeted single-gene deletion mutants in many genes that are specific to Salmonellae and not shared with close relatives, in addition to approximately 100 mutants in shared genes that served as controls, for motility phenotypes on plates [15] (Figure 1). We identified 160 mutants with altered motility. Of these mutants, only 29 were previously connected to motility either in Salmonella or E. coli. We divided the resulting set of mutants with motility phenotypes into four categories: (a) Mutants with defects in both types of motility; (b) Mutants with reduced swimming motility, swarming motility is unaffected; (c) Mutants reduced swarming motility only, swimming is unaffected; (d) Mutants that were hypermotile (Figure 2).


Identification of novel factors involved in modulating motility of Salmonella enterica serotype typhimurium.

Bogomolnaya LM, Aldrich L, Ragoza Y, Talamantes M, Andrews KD, McClelland M, Andrews-Polymenis HL - PLoS ONE (2014)

Motility scores of single deletion mutants of Salmonella Typhimurium ATCC14028 on swimming (A) and swarming (B) agar.Stationary phase cultures were transferred to motility agar, and motility was scored after 3.5 and 5 hours of incubation at 37°C on swimming or swarming plates, respectively. Motility was scored on the scale from 0 to 10 with wild type motility equal to 5. Data are presented as average swimming or swarming score from experiments with triplicate samples, performed on three independent occasions. Dots located outside of shaded area indicate scores greater than 5.25 and lower than 1.25 to define mutants with increased or reduced (<25 % of wild type) motility.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219756&req=5

pone-0111513-g001: Motility scores of single deletion mutants of Salmonella Typhimurium ATCC14028 on swimming (A) and swarming (B) agar.Stationary phase cultures were transferred to motility agar, and motility was scored after 3.5 and 5 hours of incubation at 37°C on swimming or swarming plates, respectively. Motility was scored on the scale from 0 to 10 with wild type motility equal to 5. Data are presented as average swimming or swarming score from experiments with triplicate samples, performed on three independent occasions. Dots located outside of shaded area indicate scores greater than 5.25 and lower than 1.25 to define mutants with increased or reduced (<25 % of wild type) motility.
Mentions: We have screened a library of targeted single-gene deletion mutants in many genes that are specific to Salmonellae and not shared with close relatives, in addition to approximately 100 mutants in shared genes that served as controls, for motility phenotypes on plates [15] (Figure 1). We identified 160 mutants with altered motility. Of these mutants, only 29 were previously connected to motility either in Salmonella or E. coli. We divided the resulting set of mutants with motility phenotypes into four categories: (a) Mutants with defects in both types of motility; (b) Mutants with reduced swimming motility, swarming motility is unaffected; (c) Mutants reduced swarming motility only, swimming is unaffected; (d) Mutants that were hypermotile (Figure 2).

Bottom Line: Mutants in 130 genes, not previously known to be involved in motility, had altered movement of at least one type, 9 mutants were severely impaired for both types of motility, while 33 mutants appeared defective on swimming motility plates but not swarming motility plates, and 49 mutants had reduced ability to move on swarming agar but not swimming agar.Both mutants that appeared non-motile and hypermotile on plates were assayed for expression levels of FliC and FljB on the bacterial surface and many of them had altered levels of these proteins.The phenotypes we report are the first phenotypes ever assigned to 74 of these open reading frames, as they are annotated as 'hypothetical genes' in the Typhimurium genome.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University, Bryan, Texas, United States of America; Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Russia.

ABSTRACT
Salmonella enterica serotype Typhimurium can move through liquid using swimming motility, and across a surface by swarming motility. We generated a library of targeted deletion mutants in Salmonella Typhimurium strain ATCC14028, primarily in genes specific to Salmonella, that we have previously described. In the work presented here, we screened each individual mutant from this library for the ability to move away from the site of inoculation on swimming and swarming motility agar. Mutants in genes previously described as important for motility, such as flgF, motA, cheY are do not move away from the site of inoculation on plates in our screens, validating our approach. Mutants in 130 genes, not previously known to be involved in motility, had altered movement of at least one type, 9 mutants were severely impaired for both types of motility, while 33 mutants appeared defective on swimming motility plates but not swarming motility plates, and 49 mutants had reduced ability to move on swarming agar but not swimming agar. Finally, 39 mutants were determined to be hypermotile in at least one of the types of motility tested. Both mutants that appeared non-motile and hypermotile on plates were assayed for expression levels of FliC and FljB on the bacterial surface and many of them had altered levels of these proteins. The phenotypes we report are the first phenotypes ever assigned to 74 of these open reading frames, as they are annotated as 'hypothetical genes' in the Typhimurium genome.

Show MeSH
Related in: MedlinePlus