Loss of adherence ability to human gingival epithelial cells in S-layer protein-deficient mutants of Tannerella forsythensis
Loss of adherence ability to human gingival epithelial cells in S-layer protein-deficient mutants of Tannerella forsythensis
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DOI：
10.1099/mic.0.29275-0
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发表时间：
2007-03-01
期刊：
影响因子：
2.8
通讯作者：
Yoshimura, Fuminobu
中科院分区：
文献类型：
--
作者：
Sakakibara, Junpei;Nagano, Keiji;Yoshimura, Fuminobu
福氏唐纳氏菌是牙周病的重要病原菌之一，具有典型的表层(S层)，由外膜外规则排列的亚单位组成。连翘的S层被认为与宿主细胞的血凝活性、黏附和侵袭有关，但其确切的功能尚不清楚。编码连翘主要S层蛋白(2 30和2 70 kDa)的ORF分别为TfsA、TfsA和TfsB，与最近报道的名称相同[Lee，S.-W.，Sabet，M.，Um，H.S.，Yang，L.，Kim，H.C.&朱，W.(2006年)。基因371、102-111]。为了验证S层蛋白的功能，通过聚合酶链式反应的重叠方法，成功地构建了三个tfsA和/或tfsB缺失的突变体。S-层蛋白在双突变体中完全丢失。单个缺失突变体似乎丢失了230和270 kDa蛋白中的一种。薄片显微镜清楚地显示，230和270 kDa的蛋白质组成了S层。虽然S层蛋白可能与血凝活性关系不大，但这些蛋白与人牙龈上皮细胞(Ca9-22)和KB细胞的黏附作用很强。这些结果表明，连翘S层蛋白在包括牙周病在内的口腔感染的起始阶段起重要作用。
Tannerella forsythensis, one of the important pathogens in periodontal disease, has a typical surface layer (S-layer) consisting of regularly arrayed subunits outside the outer membrane. The S-layer in T forsythensis is suggested to be associated with haemagglutinating activity, adhesion and invasion of host cells; however, its precise functions have been unknown. ORFs encoding the major S-layer proteins (230 and 270 kDa) of T forsythensis ATCC 43037, tfsA and tfsB, respectively, following the names in a recent report [Lee, S.-W., Sabet, M., Um, H. S., Yang, L., Kim, H. C. & Zhu, W. (2006). Gene 371, 102-111] were determined. To verify the function of the S-layer proteins, three mutants with tfsA, tfsB, or both deleted were successfully constructed by a PCR-based overlapping method. S-layer proteins were completely lost in the double mutant. The single-deletion mutants appeared to lose one of the 230 and 270 kDa proteins. Thin-section microscopy clearly revealed that the 230 and 270 kDa proteins composed the S-layer. Although the S-layer proteins may be weakly related to haemagglutinating activity, these proteins were highly responsible for adherence to human gingival epithelial cells (Ca9-22) and KB cells. These results suggest that the S-layer proteins in T. forsythensis play an important role in the initiation stage of oral infection including periodontal disease.