Institute of Microbial Technology, Council of Scientific and Industrial Research, Molecular Biology Division, Chandigarh 160036, India.
Gene. 2013 Aug 1;525(1):116-23. doi: 10.1016/j.gene.2013.04.071. Epub 2013 May 13.
VopF, the type III effector molecule, has been implicated in the pathogenesis of non-O1, non-O139 strains of Vibrio cholerae. It is a protein of 530 amino acids, comprises of one formin homology 1-like (FH1-like) domain and three WASP homology 2 (WH2) domains. Previous works have demonstrated that WH2 domains are crucial for VopF function as a modulator of cellular actin homeostasis. Furthermore, domain deletion analysis also suggests that VopF variant constituted with only WH2 domain 3 is more efficient in restricting the growth of budding yeast than its congeners containing either only domain 1 or domain 2. Interestingly, a good degree of sequence diversity is present within each WH2 domain of VopF. In order to ascertain the importance of different amino acids in each WH2 domain, a systemic alanine scanning mutagenesis was employed. Using a yeast model system, the alanine derivatives of each amino acid of WH2 domain 1 and 3 of VopF were examined for growth restricting activity. Taken together, our mutagenesis results reveal the identification of critical residues of WH2 domain 1 and 3 of VopF.
VopF，III 型效应分子，与非 O1、非 O139 霍乱弧菌的发病机制有关。它是一种由 530 个氨基酸组成的蛋白质，包含一个formin 同源物 1 样（FH1-like）结构域和三个 Wiskott-Aldrich 综合征蛋白 2（WASP homology 2，WH2）结构域。先前的研究表明，WH2 结构域对于 VopF 作为细胞肌动蛋白动态平衡调节剂的功能至关重要。此外，结构域缺失分析还表明，仅包含 WH2 结构域 3 的 VopF 变体比其仅包含结构域 1 或结构域 2 的同系物更有效地限制出芽酵母的生长。有趣的是，VopF 的每个 WH2 结构域内都存在相当程度的序列多样性。为了确定每个 WH2 结构域中不同氨基酸的重要性，我们采用了系统的丙氨酸扫描诱变。使用酵母模型系统，研究了 VopF 的 WH2 结构域 1 和 3 中每个氨基酸的丙氨酸衍生物对生长的限制活性。总的来说，我们的诱变结果揭示了 VopF 的 WH2 结构域 1 和 3 的关键残基。
