Insertional Inactivation and Gene Complementation of Prevotella intermedia Type IX Secretion System Reveals Its Indispensable Roles in Black Pigmentation, Hemagglutination, Protease Activity of Interpain A, and Biofilm Formation
Insertional Inactivation and Gene Complementation of Prevotella intermedia Type IX Secretion System Reveals Its Indispensable Roles in Black Pigmentation, Hemagglutination, Protease Activity of Interpain A, and Biofilm Formation
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DOI：
10.1128/jb.00203-22
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发表时间：
2022-07-13
影响因子：
3.2
通讯作者：
Nakayama，Koji
中科院分区：
文献类型：
--
作者：
Naito，Mariko;Shoji，Mikio;Nakayama，Koji
中普雷沃氏菌是一种革兰氏阴性口腔厌氧细菌，常从慢性牙周炎患者的牙周袋中分离出来。近年来，该细菌在呼吸道感染和口腔感染中的作用已被揭示出来。中间媒介虫具有多种强毒因子，如由inpagene编码的半胱氨酸蛋白酶interpain A。中间芽孢杆菌的基因组携带IX型分泌系统（T9SS）基因，该基因使毒力因子能够在属于拟杆菌门的几种病原体的外膜上易位；然而，目前尚不清楚T9SS是否在这种微生物中起作用。最近，我们对P. intermedia菌株OMA14进行了靶向诱变。在这里，我们成功地获得了缺乏inpa2和T9SS成分基因porkandport的突变体。没有一个突变体表现出interpain a的蛋白酶活性，而porkandportmutants，而inpamutant，在菌落色素沉着、血凝和生物膜形成方面表现出缺陷。我们还获得了一个补充菌株，该菌株恢复了上述所有能力。这些结果表明，T9SS在P. intermedia中起作用，interpain A是T9SS的一种货运蛋白。重要意义牙周病原体如中间普雷沃氏菌的毒力因素尚未阐明。利用我们建立的程序，我们成功地产生了IX型分泌系统突变体和基因互补菌株，这些菌株可能将毒力因子转移到细菌表面。所产生的菌株清楚地表明，中间疟原虫中的T9SS对殖民地色素沉着、血凝和生物膜的形成至关重要。这些结果表明interpain A是一种T9SS货运蛋白。
Prevotella intermedia, a Gram-negative oral anaerobic bacterium, is frequently isolated from the periodontal pockets of patients with chronic periodontitis. In recent years, the involvement of the bacterium in respiratory tract infections as well as in oral infections has been revealed. P. intermedia possesses several potent virulence factors, such as cysteine proteinase interpain A encoded by theinpAgene. The genome of P. intermedia carries genes of the type IX secretion system (T9SS), which enables the translocation of virulence factors across the outer membrane in several pathogens belonging to the phylumBacteroidetes; however, it is still unclear whether the T9SS is functional in this microorganism. Recently, we performed targeted mutagenesis in the strain OMA14 of P. intermedia. Here, we successfully obtained mutants deficient ininpAand the T9SS component genesporKandporT. None of the mutants exhibited protease activity of interpain A. TheporKandporTmutants, but not theinpAmutant, showed defects in colony pigmentation, hemagglutination, and biofilm formation. We also obtained a complemented strain for theporKgene that recovered all the above abilities. These results indicate that T9SS functions in P. intermedia and that interpain A is one of the T9SS cargo proteins.IMPORTANCEThe virulence factors of periodontal pathogens such as Prevotella intermedia have not been elucidated. Using our established procedure, we succeeded in generating type IX secretion system mutants and gene complementation strains that might transfer virulence factors to the bacterial surface. The generated strains clearly indicate that T9SS in P. intermedia is essential for colonial pigmentation, hemagglutination, and biofilm formation. These results indicated that interpain A is a T9SS cargo protein.