Deletion of lipoprotein PG0717 in Porphyromonas gingivalis W83 reduces gingipain activity and alters trafficking in and response by host cells.
Deletion of lipoprotein PG0717 in Porphyromonas gingivalis W83 reduces gingipain activity and alters trafficking in and response by host cells.
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DOI：
10.1371/journal.pone.0074230
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Progulske-Fox A
中科院分区：
文献类型：
--
作者：
Reyes L;Eiler-McManis E;Rodrigues PH;Chadda AS;Wallet SM;Bélanger M;Barrett AG;Alvarez S;Akin D;Dunn WA Jr;Progulske-Fox A
牙龈卟啉单胞菌（Pg）是慢性全身性牙周炎的病原体，与促进心血管疾病有关。 Pg株W83的脂蛋白基因PG0717的表达被发现在侵袭人冠状动脉内皮细胞(HCAEC)期间短暂上调，表明该蛋白可能与毒力有关。我们表征了 pg W83 的 PG0717 缺失突变体的毒力表型。 W83Δ717粘附和侵入HCAEC的能力没有差异。然而，W83Δ717 并未观察到接种后 24 小时内化 W83 比例增加。 PG0717 的缺失还损害了 W83 篡夺 HCAEC 中自噬途径并诱导 Saos-2 肉瘤细胞中自噬的能力。与W83相比，感染W83Δ717的HCAEC还分泌显着更大量的MCP-1、IL-8、IL-6、GM-CSF以及可溶性ICAM-1、VCAM-1和E-选择素。 W83Δ717 的进一步表征表明，荚膜和脂质 A 结构均不受 PG0717 删除的影响。有趣的是，W83Δ717 的全细胞提取物和培养上清液中精氨酸 (Rgp) 和赖氨酸 (Kgp) 牙龈蛋白酶的活性均降低。 RT-PCR 显示 rgpB 转录相应减少，但 rgpA 或 kgp 没有相应减少。两种菌株的定量蛋白质组研究表明，W83Δ717 中的 RgpA 和 RgpB 以及假定的毒力因子肽基精氨酸脱亚氨酶和 Clp 蛋白酶均显着降低。我们的结果表明，PG0717 对 W83 具有多效性，影响微生物诱导的宿主反应操纵，这对微生物清除和感染控制很重要。
P. gingivalis (Pg), a causative agent of chronic generalized periodontitis, has been implicated in promoting cardiovascular disease. Expression of lipoprotein gene PG0717 of Pg strain W83 was found to be transiently upregulated during invasion of human coronary artery endothelial cells (HCAEC), suggesting this protein may be involved in virulence. We characterized the virulence phenotype of a PG0717 deletion mutant of pg W83. There were no differences in the ability of W83Δ717 to adhere and invade HCAEC. However, the increased proportion of internalized W83 at 24 hours post-inoculation was not observed with W83∆717. Deletion of PG0717 also impaired the ability of W83 to usurp the autophagic pathway in HCAEC and to induce autophagy in Saos-2 sarcoma cells. HCAEC infected with W83Δ717 also secreted significantly greater amounts of MCP-1, IL-8, IL-6, GM-CSF, and soluble ICAM-1, VCAM-1, and E-selectin when compared to W83. Further characterization of W83Δ717 revealed that neither capsule nor lipid A structure was affected by deletion of PG0717. Interestingly, the activity of both arginine (Rgp) and lysine (Kgp) gingipains was reduced in whole-cell extracts and culture supernatant of W83Δ717. RT-PCR revealed a corresponding decrease in transcription of rgpB but not rgpA or kgp. Quantitative proteome studies of the two strains revealed that both RgpA and RgpB, along with putative virulence factors peptidylarginine deiminase and Clp protease were significantly decreased in the W83Δ717. Our results suggest that PG0717 has pleiotropic effects on W83 that affect microbial induced manipulation of host responses important for microbial clearance and infection control.
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影响因子：
3.4
作者：
Coats SR;Jones JW;Do CT;Braham PH;Bainbridge BW;To TT;Goodlett DR;Ernst RK;Darveau RP
通讯作者：
Darveau RP
影响因子：
4.2
作者：
Brunner J;Wittink FR;Jonker MJ;de Jong M;Breit TM;Laine ML;de Soet JJ;Crielaard W
通讯作者：
Crielaard W
影响因子：
56.9
作者：
Guo, L;Lim, KB;Miller, SI
通讯作者：
Miller, SI
影响因子：
3.2
作者：
Chen, T;Hosogi, Y;Duncan, MJ
通讯作者：
Duncan, MJ
影响因子：
3.1
作者：
Al-Qutub, Montaser N.;Braham, Pamela H.;Darveau, Richard P.
通讯作者：
Darveau, Richard P.