Eckl Julia, Sima Siyuan, Marcus Katrin, Lindemann Claudia, Richter Klaus
Center for Integrated Protein Science at the Technische Universität München, Department Chemie, Lichtenbergstr.Garching, Germany.
Ruhr-University Bochum, Medizinisches Proteom-Center, Functional Proteomics, Universitätsstrasse, Bochum, Germany.
PLoS One. 2017 Oct 27;12(10):e0186386. doi: 10.1371/journal.pone.0186386. eCollection 2017.
Hsp90 is a molecular chaperone involved in the regulation and maturation of kinases and transcription factors. In Caenorhabditis elegans, it contributes to the development of fertility, maintenance of muscle structure, the regulation of heat-shock response and dauer state. To understand the consequences of Hsp90-depletion, we studied Hsp90 RNAi-treated nematodes by DNA microarrays and mass spectrometry. We find that upon development of phenotypes the levels of chaperones and Hsp90 cofactors are increased, while specific proteins related to the innate immune response are depleted. In microarrays, we further find many differentially expressed genes related to gonad and larval development. These genes form an expression cluster that is regulated independently from the immune response implying separate pathways of Hsp90-involvement. Using fluorescent reporter strains for the differentially expressed immune response genes skr-5, dod-24 and clec-60 we observe that their activity in intestinal tissues is influenced by Hsp90-depletion. Instead, effects on the development are evident in both gonad arms. After Hsp90-depletion, changes can be observed in early embryos and adults containing fluorescence-tagged versions of SEPA-1, CAV-1 or PUD-1, all of which are downregulated after Hsp90-depletion. Our observations identify molecular events for Hsp90-RNAi induced phenotypes during development and immune responses, which may help to separately investigate independent Hsp90-influenced processes that are relevant during the nematode's life and development.
热休克蛋白90（Hsp90）是一种分子伴侣，参与激酶和转录因子的调控与成熟。在秀丽隐杆线虫中，它有助于生育能力的发育、肌肉结构的维持、热休克反应的调节以及滞育状态的维持。为了了解Hsp90缺失的后果，我们通过DNA微阵列和质谱技术研究了经Hsp90 RNA干扰处理的线虫。我们发现，在出现表型时，伴侣蛋白和Hsp90辅助因子的水平会升高，而与先天免疫反应相关的特定蛋白质会减少。在微阵列分析中，我们进一步发现许多与性腺和幼虫发育相关的差异表达基因。这些基因形成一个表达簇，其调控独立于免疫反应，这意味着Hsp90参与的途径是分开的。使用针对差异表达的免疫反应基因skr-5、dod-24和clec-60的荧光报告菌株，我们观察到它们在肠道组织中的活性受Hsp90缺失的影响。相反，对发育的影响在两个性腺臂中都很明显。Hsp90缺失后，在含有荧光标记版本的SEPA-1、CAV-1或PUD-1的早期胚胎和成虫中可以观察到变化，所有这些在Hsp90缺失后都会下调。我们的观察结果确定了Hsp90 RNA干扰在发育和免疫反应过程中诱导表型的分子事件，这可能有助于分别研究线虫生命和发育过程中与Hsp90相关的独立影响过程。
