Porphyromonas gingivalis Tyrosine Phosphatase Php1 Promotes Community Development and Pathogenicity
Porphyromonas gingivalis Tyrosine Phosphatase Php1 Promotes Community Development and Pathogenicity
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DOI：
10.1128/mbio.02004-19
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发表时间：
2019-09-01
期刊：
影响因子：
6.4
通讯作者：
Lamont, Richard J.
中科院分区：
文献类型：
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作者：
Jung, Young-Jung;Miller, Daniel P.;Lamont, Richard J.
细菌中的蛋白质酪氨酸磷酸化在多种细胞功能中起着重要作用，包括与群落发育和毒力相关的功能。金属依赖性蛋白酪氨酸磷酸酶属于聚合酶和组氨醇磷酸酶（PHP）家族，广泛存在于革兰氏阳性菌中。在这里，我们表明，牙龈卟啉单胞菌，革兰氏阴性牙周病原体，表达PHP蛋白，Php 1，与二价金属离子依赖性酪氨酸磷酸酶活性。Php 1酪氨酸磷酸酶活性减弱的保守的组氨酸残基的突变是重要的金属离子的协调和保守的精氨酸残基，在其他细菌PHPs催化的关键残基的突变。php 1基因位于编码细菌酪氨酸（BY）激酶Ptk 1的基因的下游，Ptk 1是Php 1在体外的底物。在没有可辨别的中间磷酸形式的情况下，Php 1迅速引起Ptk 1转化为低酪氨酸磷酸化状态。牙龈卟啉单胞菌胞外多糖的生产和社区发展所需的前口腔生物膜成分戈登链球菌在营养耗尽的条件下。相比之下，Php 1的缺乏对牙龈卟啉单胞菌形成单种生物膜的能力没有影响。在体外，Php 1酶活性是抵抗链球菌分泌的代谢产物pABA和H2 O2，抑制Ltp 1，在低分子量（LMW）磷酸酪氨酸磷酸酶家族的酶的影响。Ptk 1在酪氨酸残基159和161上磷酸化Php 1，这独立地影响磷酸酶活性。在牙周病动物模型中，Php 1的缺失使牙龈卟啉单胞菌无毒力。总的来说，这些结果表明，牙龈卟啉单胞菌具有活跃的PHP和LMW酪氨酸磷酸酶，这是革兰氏阴性菌中的一种独特构型，可使牙龈卟啉单胞菌在多物种群落中维持磷酸化/去磷酸化稳态。此外，Php 1有助于致病潜力的有机体。重要性牙周病是人类最常见的感染之一，也与全身炎症条件。牙龈卟啉单胞菌的定殖和致病性受基于蛋白质酪氨酸磷酸化和去磷酸化的信号转导途径调节。在这里，我们确定和表征的酪氨酸（去）磷酸化轴的一个新的组成部分：聚合酶和组氨醇磷酸酶（PHP）家族酶。这种酪氨酸磷酸酶，命名为Php 1，是牙龈卟啉单胞菌与其他口腔细菌共同发展所必需的，在没有Php 1活性的情况下，牙龈卟啉单胞菌不能在牙周炎小鼠模型中引起疾病。这项工作为牙龈卟啉单胞菌中的蛋白酪氨酸磷酸化（去磷酸化）网络，其对异型群落的适应及其对定植和毒力的贡献提供了重要见解。
Protein-tyrosine phosphorylation in bacteria plays a significant role in multiple cellular functions, including those related to community development and virulence. Metal-dependent protein tyrosine phosphatases that belong to the polymerase and histindinol phosphatase (PHP) family are widespread in Gram-positive bacteria. Here, we show that Porphyromonas gingivalis, a Gram-negative periodontal pathogen, expresses a PHP protein, Php1, with divalent metal ion-dependent tyrosine phosphatase activity. Php1 tyrosine phosphatase activity was attenuated by mutation of conserved histidine residues that are important for the coordination of metal ions and by mutation of a conserved arginine residue, a key residue for catalysis in other bacterial PHPs. The php1 gene is located immediately downstream of the gene encoding the bacterial tyrosine (BY) kinase Ptk1, which was a substrate for Php1 in vitro. Php1 rapidly caused the conversion of Ptk1 to a state of low tyrosine phosphorylation in the absence of discernible intermediate phosphoforms. Active Php1 was required for P. gingivalis exopolysaccharide production and for community development with the antecedent oral biofilm constituent Streptococcus gordonii under nutrient-depleted conditions. In contrast, the absence of Php1 had no effect on the ability of P. gingivalis to form monospecies biofilms. In vitro, Php1 enzymatic activity was resistant to the effects of the streptococcal secreted metabolites pABA and H2O2, which inhibited Ltp1, an enzyme in the low-molecular-weight (LMW) phosphotyrosine phosphatase family. Ptk1 reciprocally phosphorylated Php1 on tyrosine residues 159 and 161, which independently impacted phosphatase activity. Loss of Php1 rendered P. gingivalis nonvirulent in an animal model of periodontal disease. Collectively, these results demonstrate that P. gingivalis possesses active PHP and LMW tyrosine phosphatases, a unique configuration in Gram-negatives which may allow P. gingivalis to maintain phosphorylation/dephosphorylation homeostasis in multispecies communities. Moreover, Php1 contributes to the pathogenic potential of the organism.IMPORTANCE Periodontal diseases are among the most common infections of humans and are also associated with systemic inflammatory conditions. Colonization and pathogenicity of P. gingivalis are regulated by signal transduction pathways based on protein tyrosine phosphorylation and dephosphorylation. Here, we identify and characterize a novel component of the tyrosine (de)phosphorylation axis: a polymerase and histindinol phosphatase (PHP) family enzyme. This tyrosine phosphatase, designated Php1, was required for P. gingivalis community development with other oral bacteria, and in the absence of Php1 activity P. gingivalis was unable to cause disease in a mouse model of periodontitis. This work provides significant insights into the protein tyrosine (de)phosphorylation network in P. gingivalis, its adaptation to heterotypic communities, and its contribution to colonization and virulence.