The major inducible small heat shock protein HSP20-3 in the tardigrade Ramazzottius varieornatus forms filament-like structures and is an active chaperone

IF 3.3 3区 生物学 Q3 CELL BIOLOGY
Mohammad Al-Ansari , Taylor Fitzsimons , Wenbin Wei , Martin W. Goldberg , Takekazu Kunieda , Roy A. Quinlan
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引用次数: 0

Abstract

The tardigrade Ramazzottius varieornatus has remarkable resilience to a range of environmental stresses. In this study, we have characterised two members of the small heat shock protein (sHSP) family in R. varieornatus, HSP20–3 and HSP20–6. These are the most highly upregulated sHSPs in response to a 24 h heat shock at 35 0C of adult tardigrades with HSP20–3 being one of the most highly upregulated gene in the whole transcriptome. Both R. varieornatus sHSPs and the human sHSP, CRYAB (HSPB5), were produced recombinantly for comparative structure-function studies. HSP20–3 exhibited a superior chaperone activity than human CRYAB in a heat-induced protein aggregation assay. Both tardigrade sHSPs also formed larger oligomers than CRYAB as assessed by size exclusion chromatography and transmission electron microscopy of negatively stained samples. Whilst both HSP20–3 and HSP20–6 formed particles that were variable in size and larger than the particles formed by CRYAB, only HSP20–3 formed filament-like structures. The particles and filament-like structures formed by HSP20–3 appear inter-related as the filament-like structures often had particles located at their ends. Sequence analyses identified two unique features; an insertion in the middle region of the N-terminal domain (NTD) and preceding the critical-sequence identified in CRYAB, as well as a repeated QNTN-motif located in the C-terminal domain of HSP20–3. The NTD insertion is expected to affect protein-protein interactions and subunit oligomerisation. Removal of the repeated QNTN-motif abolished HSP20–3 chaperone activity and also affected the assembly of the filament-like structures. We discuss the potential contribution of HSP20–3 to protein condensate formation.

在沙蜥(Ramazzottius varieornatus)中,主要的诱导性小型热休克蛋白 HSP20-3 形成丝状结构,是一种活性伴侣蛋白
变角龙蜥对一系列环境压力具有非凡的适应能力。在这项研究中,我们鉴定了R. varieornatus体内小型热休克蛋白(sHSP)家族的两个成员,即HSP20-3和HSP20-6。这两种小热休克蛋白是成年沙蜥在 35 摄氏度下受到 24 小时热休克后上调幅度最大的小热休克蛋白,其中 HSP20-3 是整个转录组中上调幅度最大的基因之一。为了进行结构-功能比较研究,我们重组生产了变角龙 sHSP 和人类 sHSP CRYAB(HSPB5)。在热诱导蛋白质聚集试验中,HSP20-3 表现出比人类 CRYAB 更强的伴侣活性。通过尺寸排阻色谱法和阴性染色样品的透射电子显微镜评估,这两种短尾龙 sHSP 形成的寡聚体也比 CRYAB 大。虽然 HSP20-3 和 HSP20-6 形成的颗粒大小不一,而且比 CRYAB 形成的颗粒更大,但只有 HSP20-3 形成了丝状结构。HSP20-3 形成的颗粒和丝状结构似乎相互关联,因为丝状结构的末端往往有颗粒。序列分析发现了两个独特的特征:位于 N 端结构域(NTD)中间区域、CRYAB 中发现的临界序列之前的插入物,以及位于 HSP20-3 C 端结构域的重复 QNTN-motif。NTD 插入预计会影响蛋白质与蛋白质之间的相互作用以及亚基的寡聚化。去除重复的 QNTN-motif后,HSP20-3的伴侣蛋白活性消失,丝状结构的组装也受到影响。我们讨论了 HSP20-3 对蛋白质凝聚物形成的潜在贡献。
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来源期刊
Cell Stress & Chaperones
Cell Stress & Chaperones 生物-细胞生物学
CiteScore
7.60
自引率
2.60%
发文量
59
审稿时长
6-12 weeks
期刊介绍: Cell Stress and Chaperones is an integrative journal that bridges the gap between laboratory model systems and natural populations. The journal captures the eclectic spirit of the cellular stress response field in a single, concentrated source of current information. Major emphasis is placed on the effects of climate change on individual species in the natural environment and their capacity to adapt. This emphasis expands our focus on stress biology and medicine by linking climate change effects to research on cellular stress responses of animals, micro-organisms and plants.
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