In silico analysis of usher encoding genes in Klebsiella pneumoniae and characterization of their role in adhesion and colonization.
In silico analysis of usher encoding genes in Klebsiella pneumoniae and characterization of their role in adhesion and colonization.
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DOI：
10.1371/journal.pone.0116215
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Forestier C
中科院分区：
文献类型：
--
作者：
Khater F;Balestrino D;Charbonnel N;Dufayard JF;Brisse S;Forestier C
伴侣/聚合体(CU)组装途径被广泛的肠杆菌科利用来组装被称为菌毛或菌毛的粘附性表面结构，这些结构在细菌-宿主细胞相互作用中发挥作用。计算机分析表明，肺炎克雷伯菌LM21基因组含有8个CU基因座，分别属于γκп和ϭ簇。在这些操作子中，只有两个对应于前面描述的操作子，即类型1和类型3-编码操作子。构建了肺炎克雷伯菌LM21的等位基因引座子缺失突变体，并研究了其在动植物细胞黏附、生物膜形成和小鼠肠道定植中的作用。3型菌毛引入缺失突变体在所有检测中都受到损害，而1型菌毛引入缺失突变体仅在与植物细胞的粘附性和肠道定植方面表现出减弱。无论是单独接种还是在联合接种实验中，LM21ΔkpjC突变体在黏附拟南芥细胞和在小鼠肠道定植的能力都受到了损害。LM21kpgC的缺失导致生物膜的形成、与动物细胞的粘附性和小鼠肠道定植的显著减少。LM2 1∆kpaC和LM2 1∆kpeC突变体分别在生物膜形成和对拟南芥细胞的黏附能力方面受到抑制。LM2 1∆kpbC和Lm2 1∆kpdC突变体在体外和体内均未观察到明显的效应。肺炎克雷伯菌基因组中CU基因座的多样性及其特有的黏附模式可能反映了该细菌在其不同生态位中对不同底物的黏附能力。
Chaperone/usher (CU) assembly pathway is used by a wide range of Enterobacteriaceae to assemble adhesive surface structures called pili or fimbriae that play a role in bacteria-host cell interactions. In silico analysis revealed that the genome of Klebsiella pneumoniae LM21 harbors eight chromosomal CU loci belonging to γκп and ϭ clusters. Of these, only two correspond to previously described operons, namely type 1 and type 3-encoding operons. Isogenic usher deletion mutants of K. pneumoniae LM21 were constructed for each locus and their role in adhesion to animal (Intestine 407) and plant (Arabidopsis thaliana) cells, biofilm formation and murine intestinal colonization was investigated. Type 3 pili usher deleted mutant was impaired in all assays, whereas type 1 pili usher deleted mutant only showed attenuation in adhesion to plant cells and in intestinal colonization. The LM21ΔkpjC mutant was impaired in its capacity to adhere to Arabidopsis cells and to colonize the murine intestine, either alone or in co-inoculation experiments. Deletion of LM21kpgC induced a significant decrease in biofilm formation, in adhesion to animal cells and in colonization of the mice intestine. The LM21∆kpaC and LM21∆kpeC mutants were only attenuated in biofilm formation and the adhesion abilities to Arabidopsis cells, respectively. No clear in vitro or in vivo effect was observed for LM21∆kpbC and LM21∆kpdC mutants. The multiplicity of CU loci in K. pneumoniae genome and their specific adhesion pattern probably reflect the ability of the bacteria to adhere to different substrates in its diverse ecological niches.
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作者：
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