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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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Related in: MedlinePlus

Reduced motility on semi-solid media in some mutants correlated with reduced expression of flagellin on the bacterial surface.Flagellins were sheared from bacterial surface, analyzed by Western blotting with antibodies to FliC (black bars) and FljB (grey bars) and blots were quantified by densitometry using Quantity One software. Surface expression of flagellins was normalized to the level on the surface of the wild type strain. Samples for each mutant were prepared in three independent experiments.
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pone-0111513-g003: Reduced motility on semi-solid media in some mutants correlated with reduced expression of flagellin on the bacterial surface.Flagellins were sheared from bacterial surface, analyzed by Western blotting with antibodies to FliC (black bars) and FljB (grey bars) and blots were quantified by densitometry using Quantity One software. Surface expression of flagellins was normalized to the level on the surface of the wild type strain. Samples for each mutant were prepared in three independent experiments.

Mentions: Mutants in flgF, flgG, and rfbK, genes known to be important for motility [14], [22], were both non-motile and displayed less flagellins on the bacterial surface in our screen. For other mutants however, (motB, cheY) the presence of filaments is known not to be sufficient for the ability of these mutants to move from the site of inoculation on motility plates. Our mutants in motB and cheY displayed wild type levels of flagellins on the cell surface yet were unable to spread from the site of inoculation as previously reported in the literature [23], [24] (Figure 3).


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)

Reduced motility on semi-solid media in some mutants correlated with reduced expression of flagellin on the bacterial surface.Flagellins were sheared from bacterial surface, analyzed by Western blotting with antibodies to FliC (black bars) and FljB (grey bars) and blots were quantified by densitometry using Quantity One software. Surface expression of flagellins was normalized to the level on the surface of the wild type strain. Samples for each mutant were prepared in three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0111513-g003: Reduced motility on semi-solid media in some mutants correlated with reduced expression of flagellin on the bacterial surface.Flagellins were sheared from bacterial surface, analyzed by Western blotting with antibodies to FliC (black bars) and FljB (grey bars) and blots were quantified by densitometry using Quantity One software. Surface expression of flagellins was normalized to the level on the surface of the wild type strain. Samples for each mutant were prepared in three independent experiments.
Mentions: Mutants in flgF, flgG, and rfbK, genes known to be important for motility [14], [22], were both non-motile and displayed less flagellins on the bacterial surface in our screen. For other mutants however, (motB, cheY) the presence of filaments is known not to be sufficient for the ability of these mutants to move from the site of inoculation on motility plates. Our mutants in motB and cheY displayed wild type levels of flagellins on the cell surface yet were unable to spread from the site of inoculation as previously reported in the literature [23], [24] (Figure 3).

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