Fujiwara Nagatoshi, Ohara Naoya, Ogawa Midori, Maeda Shinji, Naka Takashi, Taniguchi Hatsumi, Yamamoto Saburo, Ayata Minoru
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.
Department of Oral Microbiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama City, Okayama, Japan.
PLoS One. 2015 May 13;10(5):e0126813. doi: 10.1371/journal.pone.0126813. eCollection 2015.
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.
耻垢分枝杆菌已被广泛用作分枝杆菌感染模型。与耻垢分枝杆菌mc(2)155菌株不同,耻垢分枝杆菌J15cs菌株具有在小鼠巨噬细胞中存活一周的优势。在我们之前的报告中,我们阐明J15cs菌株的细胞壁中缺失了非极性糖脂(GPLs),这可能会影响其形态以及在宿主细胞中的存活。在本研究中,鉴定出了导致J15cs菌株中GPL缺失的基因。mps1-2基因(MSMEG_0400 - 0402)与GPL生物合成相关。与mc(2)155菌株相比,J15cs菌株的mps1基因中缺失了18个碱基对。mps1互补的J15cs突变体恢复了GPL的表达。尽管J15cs菌株产生粗糙干燥的菌落,但这种mps1互补菌株的菌落形态像mc(2)155菌株一样光滑。mps1互补的J15cs突变体的长度因GPL的表达而缩短。此外,GPL恢复的J15cs突变体在小鼠巨噬细胞系J774.1细胞中存活的时间不如亲本J15cs菌株长。这些结果直接证明J15cs菌株中GPL的缺失会影响细菌大小、形态以及在宿主细胞中的存活。