Mir Dilawar Ahmad, Balasubramaniam Boopathi, VenkataKrishna Lappasi Mohanram, Muthubharathi Balasubramanian Chellammal, Balamurugan Krishnaswamy
Department of Biotechnology, Alagappa University, Karaikudi, Tamil Nadu, India.
Genes Immun. 2021 Jun;22(2):75-92. doi: 10.1038/s41435-021-00132-w. Epub 2021 May 14.
Bacterial effector molecules are crucial infectious agents that can cause pathogenesis. In the present study, the pathogenesis of toxic Salmonella enterica serovar Typhi (S. Typhi) proteins on the model host Caenorhabditis elegans was investigated by exploring the host's regulatory proteins during infection through the quantitative proteomics approach. Extracted host proteins were analyzed using two-dimensional gel electrophoresis (2D-GE) and differentially regulated proteins were identified using MALDI TOF/TOF/MS analysis. Of the 150 regulated proteins identified, 95 were downregulated while 55 were upregulated. The interaction network of regulated proteins was predicted using the STRING tool. Most downregulated proteins were involved in muscle contraction, locomotion, energy hydrolysis, lipid synthesis, serine/threonine kinase activity, oxidoreductase activity, and protein unfolding. Upregulated proteins were involved in oxidative stress pathways. Hence, cellular stress generated by S. Typhi proteins in the model host was determined using lipid peroxidation as well as oxidant and antioxidant assays. In addition, candidate proteins identified via extract analysis were validated by western blotting, and the roles of several crucial molecules were analyzed in vivo using transgenic strains (myo-2 and col-19) and mutant (ogt-1) of C. elegans. To the best of our knowledge, this is the first study to report protein regulation in host C. elegans exposed to toxic S. Typhi proteins. It highlights the significance of p38 MAPK and JNK immune pathways.
细菌效应分子是可导致发病机制的关键感染因子。在本研究中,通过定量蛋白质组学方法探索感染过程中宿主的调节蛋白,研究了有毒肠炎沙门氏菌伤寒血清型(伤寒杆菌)蛋白在模式宿主秀丽隐杆线虫上的发病机制。使用二维凝胶电泳(2D-GE)分析提取的宿主蛋白,并使用基质辅助激光解吸电离飞行时间/飞行时间串联质谱(MALDI TOF/TOF/MS)分析鉴定差异调节蛋白。在鉴定出的150种调节蛋白中,95种下调,55种上调。使用STRING工具预测调节蛋白的相互作用网络。大多数下调蛋白参与肌肉收缩、运动、能量水解、脂质合成、丝氨酸/苏氨酸激酶活性、氧化还原酶活性和蛋白质解折叠。上调蛋白参与氧化应激途径。因此,使用脂质过氧化以及氧化剂和抗氧化剂测定法确定了伤寒杆菌蛋白在模式宿主中产生的细胞应激。此外,通过提取物分析鉴定的候选蛋白通过蛋白质免疫印迹法进行验证,并使用秀丽隐杆线虫的转基因菌株(myo-2和col-19)和突变体(ogt-1)在体内分析了几种关键分子的作用。据我们所知,这是第一项报道暴露于有毒伤寒杆菌蛋白的宿主秀丽隐杆线虫中蛋白质调节的研究。它突出了p38丝裂原活化蛋白激酶和JNK免疫途径的重要性。