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Glycopeptidolipid of Mycobacterium smegmatis J15cs Affects Morphology and Survival in Host Cells.

Fujiwara N, Ohara N, Ogawa M, Maeda S, Naka T, Taniguchi H, Yamamoto S, Ayata M - PLoS ONE (2015)

Bottom Line: Mycobacterium smegmatis has been widely used as a mycobacterial infection model.The mps1-complemented J15cs mutant restored the expression of GPLs.The length in the mps1-complemented J15cs mutant was shortened by the expression of GPLs.

View Article: PubMed Central - PubMed

Affiliation: Department of Food and Nutrition, Faculty of Contemporary Human Life Science, Tezukayama University, Nara City, Nara, Japan; Department of Bacteriology, Osaka City University Graduate School of Medicine, Osaka City, Osaka, Japan.

ABSTRACT
Mycobacterium smegmatis has been widely used as a mycobacterial infection model. Unlike the M. smegmatis mc(2)155 strain, M. smegmatis J15cs strain has the advantage of surviving for one week in murine macrophages. In our previous report, we clarified that the J15cs strain has deleted apolar glycopeptidolipids (GPLs) in the cell wall, which may affect its morphology and survival in host cells. In this study, the gene causing the GPL deletion in the J15cs strain was identified. The mps1-2 gene (MSMEG_0400-0402) correlated with GPL biosynthesis. The J15cs strain had 18 bps deleted in the mps1 gene compared to that of the mc(2)155 strain. The mps1-complemented J15cs mutant restored the expression of GPLs. Although the J15cs strain produces a rough and dry colony, the colony morphology of this mps1-complement was smooth like the mc(2)155 strain. The length in the mps1-complemented J15cs mutant was shortened by the expression of GPLs. In addition, the GPL-restored J15cs mutant did not survive as long as the parent J15cs strain in the murine macrophage cell line J774.1 cells. The results are direct evidence that the deletion of GPLs in the J15cs strain affects bacterial size, morphology, and survival in host cells.

No MeSH data available.


Related in: MedlinePlus

Change of morphology due to expression of GPLs.The colony morphology was observed by an optical microscope after culturing. The J15cs strain formed rough, dry, and irregular colonies. The mc2155 strain and the mps1-complemented J15cs mutant colonies were smoothly curved and wet (a). Scanning electron microscopy (b) and a comparison of bacterial cell length (c) are shown. The bacteria were grown on nutrient agar plates for 10 days at 37°C, collected, and filtered through a 5 μm membrane to remove aggregates. The average bacterial cell length was 2.11±0.46 μm, the mc2155 strain; 5.70±1.65, the J15cs strain; 2.18±0.62 μm, the mps1-complemented J15cs mutant. The data are means±standard deviations (SD) for 20 bacteria. The separate experiments were done in duplicate. *, p<0.001.
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pone.0126813.g005: Change of morphology due to expression of GPLs.The colony morphology was observed by an optical microscope after culturing. The J15cs strain formed rough, dry, and irregular colonies. The mc2155 strain and the mps1-complemented J15cs mutant colonies were smoothly curved and wet (a). Scanning electron microscopy (b) and a comparison of bacterial cell length (c) are shown. The bacteria were grown on nutrient agar plates for 10 days at 37°C, collected, and filtered through a 5 μm membrane to remove aggregates. The average bacterial cell length was 2.11±0.46 μm, the mc2155 strain; 5.70±1.65, the J15cs strain; 2.18±0.62 μm, the mps1-complemented J15cs mutant. The data are means±standard deviations (SD) for 20 bacteria. The separate experiments were done in duplicate. *, p<0.001.

Mentions: After being cultured on nutrient agar for 10 days at 37°C, the J15cs strain formed rough, dry, and irregular colonies. In contrast, the colonies of the mc2155 strain were smoothly curved and wet. Thus, the morphology of the colonies was strain-specific. The J15cs strain carrying the pNN2 vector showed the same morphology as the parent J15cs strain, and the mps1-complemented J15cs mutant formed smoothly curved and wet colonies like the mc2155 strain (Fig 5A). The hydrophobicity of the mps1-complemented J15cs mutant was reduced, compared to the parent J15cs strain. The SEM images showed that the bacterial length of the J15cs strain was longer than that of the mc2155 strain, and the mps1-complemented J15cs mutant was significantly shorter than the parent J15cs strain. There was no significant difference in their widths (Fig 5B and 5C). The restoration of GPLs in the mps1-complemented J15cs mutant may modify the cell wall construction. As a result, the colony morphology and bacterial size were changed to those of the mc2155 strain. It was concluded that the existence of GPLs affected bacterial size and morphology.


Glycopeptidolipid of Mycobacterium smegmatis J15cs Affects Morphology and Survival in Host Cells.

Fujiwara N, Ohara N, Ogawa M, Maeda S, Naka T, Taniguchi H, Yamamoto S, Ayata M - PLoS ONE (2015)

Change of morphology due to expression of GPLs.The colony morphology was observed by an optical microscope after culturing. The J15cs strain formed rough, dry, and irregular colonies. The mc2155 strain and the mps1-complemented J15cs mutant colonies were smoothly curved and wet (a). Scanning electron microscopy (b) and a comparison of bacterial cell length (c) are shown. The bacteria were grown on nutrient agar plates for 10 days at 37°C, collected, and filtered through a 5 μm membrane to remove aggregates. The average bacterial cell length was 2.11±0.46 μm, the mc2155 strain; 5.70±1.65, the J15cs strain; 2.18±0.62 μm, the mps1-complemented J15cs mutant. The data are means±standard deviations (SD) for 20 bacteria. The separate experiments were done in duplicate. *, p<0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4430512&req=5

pone.0126813.g005: Change of morphology due to expression of GPLs.The colony morphology was observed by an optical microscope after culturing. The J15cs strain formed rough, dry, and irregular colonies. The mc2155 strain and the mps1-complemented J15cs mutant colonies were smoothly curved and wet (a). Scanning electron microscopy (b) and a comparison of bacterial cell length (c) are shown. The bacteria were grown on nutrient agar plates for 10 days at 37°C, collected, and filtered through a 5 μm membrane to remove aggregates. The average bacterial cell length was 2.11±0.46 μm, the mc2155 strain; 5.70±1.65, the J15cs strain; 2.18±0.62 μm, the mps1-complemented J15cs mutant. The data are means±standard deviations (SD) for 20 bacteria. The separate experiments were done in duplicate. *, p<0.001.
Mentions: After being cultured on nutrient agar for 10 days at 37°C, the J15cs strain formed rough, dry, and irregular colonies. In contrast, the colonies of the mc2155 strain were smoothly curved and wet. Thus, the morphology of the colonies was strain-specific. The J15cs strain carrying the pNN2 vector showed the same morphology as the parent J15cs strain, and the mps1-complemented J15cs mutant formed smoothly curved and wet colonies like the mc2155 strain (Fig 5A). The hydrophobicity of the mps1-complemented J15cs mutant was reduced, compared to the parent J15cs strain. The SEM images showed that the bacterial length of the J15cs strain was longer than that of the mc2155 strain, and the mps1-complemented J15cs mutant was significantly shorter than the parent J15cs strain. There was no significant difference in their widths (Fig 5B and 5C). The restoration of GPLs in the mps1-complemented J15cs mutant may modify the cell wall construction. As a result, the colony morphology and bacterial size were changed to those of the mc2155 strain. It was concluded that the existence of GPLs affected bacterial size and morphology.

Bottom Line: Mycobacterium smegmatis has been widely used as a mycobacterial infection model.The mps1-complemented J15cs mutant restored the expression of GPLs.The length in the mps1-complemented J15cs mutant was shortened by the expression of GPLs.

View Article: PubMed Central - PubMed

Affiliation: Department of Food and Nutrition, Faculty of Contemporary Human Life Science, Tezukayama University, Nara City, Nara, Japan; Department of Bacteriology, Osaka City University Graduate School of Medicine, Osaka City, Osaka, Japan.

ABSTRACT
Mycobacterium smegmatis has been widely used as a mycobacterial infection model. Unlike the M. smegmatis mc(2)155 strain, M. smegmatis J15cs strain has the advantage of surviving for one week in murine macrophages. In our previous report, we clarified that the J15cs strain has deleted apolar glycopeptidolipids (GPLs) in the cell wall, which may affect its morphology and survival in host cells. In this study, the gene causing the GPL deletion in the J15cs strain was identified. The mps1-2 gene (MSMEG_0400-0402) correlated with GPL biosynthesis. The J15cs strain had 18 bps deleted in the mps1 gene compared to that of the mc(2)155 strain. The mps1-complemented J15cs mutant restored the expression of GPLs. Although the J15cs strain produces a rough and dry colony, the colony morphology of this mps1-complement was smooth like the mc(2)155 strain. The length in the mps1-complemented J15cs mutant was shortened by the expression of GPLs. In addition, the GPL-restored J15cs mutant did not survive as long as the parent J15cs strain in the murine macrophage cell line J774.1 cells. The results are direct evidence that the deletion of GPLs in the J15cs strain affects bacterial size, morphology, and survival in host cells.

No MeSH data available.


Related in: MedlinePlus