Impact of protein domains on PE_PGRS30 polar localization in Mycobacteria.
Impact of protein domains on PE_PGRS30 polar localization in Mycobacteria.
复制标题
蛋白质结构域对分枝杆菌中PE_PGRS30极性定位的影响。
DOI：
10.1371/journal.pone.0112482
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Delogu G
中科院分区：
文献类型：
--
作者：
De Maio F;Maulucci G;Minerva M;Anoosheh S;Palucci I;Iantomasi R;Palmieri V;Camassa S;Sali M;Sanguinetti M;Bitter W;Manganelli R;De Spirito M;Delogu G
PE_PGRS蛋白是结核分枝杆菌复合体和许多其它致病性分枝杆菌所特有的。PE_PGRS30是M.结核分枝杆菌（Mtb）具有三个主要结构域，即N-末端PE结构域、重复PGRS结构域和独特的C-末端结构域。为了研究这些结构域的作用，我们表达了GFP标记的PE_PGRS30蛋白和一系列其功能缺失突变体在不同的分枝杆菌物种（结核分枝杆菌，牛分枝杆菌BCG和耻垢分枝杆菌），并分析了蛋白质定位的共聚焦显微镜。我们发现PE_PGRS30定位于Mtb和M. bovis BCG，而M.并且该蛋白的PGRS结构域强烈有助于Mtb中的蛋白细胞定位。免疫荧光研究进一步表明，PE_PGRS 30的独特C-末端结构域在表面上不可用，除非PGRS结构域缺失。免疫印迹表明，PGRS结构域是维持蛋白质与不溶性细胞组分强烈相关所必需的。这些结果表明，PGRS结构域的重复GGA-GGN重复序列包含有助于蛋白质细胞定位的特定序列，并且极性定位可能是PE_PGRS30依赖性毒力机制中的关键步骤。
PE_PGRS proteins are unique to the Mycobacterium tuberculosis complex and a number of other pathogenic mycobacteria. PE_PGRS30, which is required for the full virulence of M. tuberculosis (Mtb), has three main domains, i.e. an N-terminal PE domain, repetitive PGRS domain and the unique C-terminal domain. To investigate the role of these domains, we expressed a GFP-tagged PE_PGRS30 protein and a series of its functional deletion mutants in different mycobacterial species (Mtb, Mycobacterium bovis BCG and Mycobacterium smegmatis) and analysed protein localization by confocal microscopy. We show that PE_PGRS30 localizes at the mycobacterial cell poles in Mtb and M. bovis BCG but not in M. smegmatis and that the PGRS domain of the protein strongly contributes to protein cellular localization in Mtb. Immunofluorescence studies further showed that the unique C-terminal domain of PE_PGRS30 is not available on the surface, except when the PGRS domain is missing. Immunoblot demonstrated that the PGRS domain is required to maintain the protein strongly associated with the non-soluble cellular fraction. These results suggest that the repetitive GGA-GGN repeats of the PGRS domain contain specific sequences that contribute to protein cellular localization and that polar localization might be a key step in the PE_PGRS30-dependent virulence mechanism.
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DOI：
10.1073/pnas.1119453109
发表时间：
2012-07-10
影响因子：
11.1
作者：
Daleke, Maria H.;Ummels, Roy;Bitter, Wilbert
通讯作者：
Bitter, Wilbert
影响因子：
2.1
作者：
Chatrath, Shweta;Gupta, Vineet Kumar;Garg, Lalit C.
通讯作者：
Garg, Lalit C.
影响因子：
6.7
作者：
Carlsson, Fredric;Joshi, Shilpa A.;Rangell, Linda;Brown, Eric J.
通讯作者：
Brown, Eric J.
影响因子：
3.7
作者：
Cascioferro A;Daleke MH;Ventura M;Donà V;Delogu G;Palù G;Bitter W;Manganelli R
通讯作者：
Manganelli R
影响因子：
5.8
作者：
Delogu, Giovanni;Sanguinetti, Maurizio;Fadda, Giovanni
通讯作者：
Fadda, Giovanni