The arginine-ornithine antiporter ArcD contributes to biological fitness of Streptococcus suis.
The arginine-ornithine antiporter ArcD contributes to biological fitness of Streptococcus suis.
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DOI：
10.3389/fcimb.2014.00107
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发表时间：
2014
影响因子：
5.7
通讯作者：
Goethe R
中科院分区：
文献类型：
--
作者：
Fulde M;Willenborg J;Huber C;Hitzmann A;Willms D;Seitz M;Eisenreich W;Valentin-Weigand P;Goethe R
精氨酸-鸟氨酸反向转运蛋白（ArcD）是精氨酸脱亚胺酶系统（ADS）的一部分，ADS是一种在多种不同细菌物种（包括猪人畜共患病病原体猪链球菌）中发现的分解代谢能量提供途径。最近的研究表明，ADS在S.特别是在宿主细胞中的存活。然而，精氨酸和精氨酸转运介导的ArcD的贡献，还有待澄清。本研究以[U-13 C6]精氨酸为示踪分子，通过实验证明，S.猪是精氨酸营养缺陷型，细菌生长依赖于细胞外精氨酸的摄取。为了进一步研究ArcD在精氨酸代谢中的作用，我们产生了一种arcD特异性突变株，并将其生长特征与野生型（WT）菌株（一种强毒血清型2菌株）进行了比较。突变株显示出显着降低的生长在补充有精氨酸的化学成分确定的培养基相比，野生型菌株，表明ArcD促进精氨酸摄取。为了进一步评估ArcD的体内相关性，我们研究了上皮细胞培养感染模型中arcD突变株的细胞内细菌存活。突变株基本上减毒，其降低的细胞内存活率与较低的中和酸化环境的能力相关。基于这些结果，我们提出ArcD通过其作为精氨酸-鸟氨酸反向转运蛋白的功能，对于供应作为ADS底物的精氨酸是重要的，从而有助于S.猪在宿主中。
The arginine-ornithine antiporter (ArcD) is part of the Arginine Deiminase System (ADS), a catabolic, energy-providing pathway found in a variety of different bacterial species, including the porcine zoonotic pathogen Streptococcus suis. The ADS has recently been shown to play a role in the pathogenicity of S. suis, in particular in its survival in host cells. The contribution of arginine and arginine transport mediated by ArcD, however, has yet to be clarified. In the present study, we showed by experiments using [U-13C6]arginine as a tracer molecule that S. suis is auxotrophic for arginine and that bacterial growth depends on the uptake of extracellular arginine. To further study the role of ArcD in arginine metabolism, we generated an arcD-specific mutant strain and characterized its growth compared to the wild-type (WT) strain, a virulent serotype 2 strain. The mutant strain showed a markedly reduced growth in chemically defined media supplemented with arginine when compared to the WT strain, suggesting that ArcD promotes arginine uptake. To further evaluate the in vivo relevance of ArcD, we studied the intracellular bacterial survival of the arcD mutant strain in an epithelial cell culture infection model. The mutant strain was substantially attenuated, and its reduced intracellular survival rate correlated with a lower ability to neutralize the acidified environment. Based on these results, we propose that ArcD, by its function as an arginine-ornithine antiporter, is important for supplying arginine as substrate of the ADS and, thereby, contributes to biological fitness and virulence of S. suis in the host.
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