A novel type of two-component regulatory system affecting gingipains in Porphyromonas gingivalis
A novel type of two-component regulatory system affecting gingipains in Porphyromonas gingivalis
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DOI：
10.1111/j.1348-0421.2003.tb03451.x
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发表时间：
2003-01-01
影响因子：
2.6
通讯作者：
Yoshimura, F
中科院分区：
文献类型：
--
作者：
Hasegawa, Y;Nishiyama, S;Yoshimura, F
我们调查了牙龈卟啉单胞菌W83基因组数据库中FIMS的同源物，FIMS是第一个可能控制毒力因子的双组分传感器组氨酸激酶。包括FIMs在内，我们在基因组中发现了六个可能的传感器激酶基因。从牙龈假单胞菌质粒库中克隆了编码其中一个同源物的基因，并对其进行了测序和突变体分析。通过在基因中引入一个药盒，在atcc 33277菌株中产生了两个基因中断突变体。突变体形成了无色素的菌落，这表明它们可能在产生蛋白酶方面存在缺陷，这是这种有机体的一个特征。用精氨酸和赖氨酸专一性(分别是RGP和KGP牙龈痛)来衡量，突变株的蛋白酶活性几乎是亲本菌株的一半。与亲本菌株和野生型菌株不同，大多数牙周疼痛活性在突变株的培养上清液中检测到，而不是在细胞中检测到。十二烷基硫酸钠聚丙烯酰胺凝胶电泳法和Western印迹分析证实了牙周疼痛的异常产生。这些结果有力地表明，这种新发现的双组分感受器激酶通过一种未知的机制参与了牙龈痛的成熟和外膜的适当定位。编码感受器组氨酸激酶的基因被命名为gppX，它代表了在bar下的(P)中对牙周疼痛的调节(X)和对黑色(P)的调节。牙周炎。
We surveyed the Porphyromonas gingivalis W83 genome database for homologues of FimS, the first two-component sensor histidine kinase, which could possibly control virulence factors. Including fimS, we found six putative sensor kinase genes in the genome. The gene encoding one of the homologues was cloned from a P gingivalis plasmid library, sequenced, and analyzed using its mutants. Two gene-disruption mutants were created in strain ATCC 33277 by introducing a drug cassette into the gene. The mutants formed nonpigmented colonies, indicating that they might be defective in proteinase production, a characteristic of this organism. Proteinase activities, measured as arginine- and lysine-specific (Rgp and Kgp gingipains, respectively) activities, of the mutants were almost half those of the parent strain. Unlike the parent and wildtype strains, most of the gingipain activities were detected in the culture supernatant, not in cells, of the mutants. Abnormal production of gingipains was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analyses. These results strongly suggest that this newly-discovered two-component sensor kinase is involved in maturation and proper localization of gingipains to the outer membrane through an unknown mechanism. The gene encoding the sensor histidine kinase was designated gppX, which represents regulation (X) of gingipains and black (p) under bar igmentation in (P) under bar. gingivalis.