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在集胞藻 PCC 6803 中过表达 ClpB1 可提高其对快速热冲击的耐受性。

ClpB1 overproduction in Synechocystis sp. strain PCC 6803 increases tolerance to rapid heat shock.

机构信息

School of Life Sciences and Center for Bioenergy and Photosynthesis, Arizona State University, Tempe, Arizona, USA.

出版信息

Appl Environ Microbiol. 2013 Oct;79(20):6220-7. doi: 10.1128/AEM.01661-13. Epub 2013 Aug 2.

Abstract

ClpB1 is a heat shock protein known to disaggregate large protein complexes. Constitutive, 16-fold ClpB1 overproduction in the cyanobacterium Synechocystis sp. strain PCC 6803 increased cell survival by 20-fold when cultures were heated quickly (1°C/s) to 50°C and delayed cell death by an average of 3 min during incubation at high temperatures (>46°C). Cooverexpression of ClpB1 and another heat shock protein, DnaK2, further increased cell survival. According to immunocytochemistry results, ClpB1 is dispersed throughout the cytoplasm but is concentrated in specific areas and is more prevalent near thylakoid membranes. However, ClpB1 overproduction does not lead to a change in the morphology, chlorophyll content, or photosystem ratio. Whereas electron microscopy demonstrated that apparent protein aggregation occurred after heat treatment in the control strain, protein aggregate size was maintained in the ClpB1 overexpresser. Constitutive ClpB1 overproduction allows an earlier response to heat shock and protects from rapid heating of cultures.

摘要

ClpB1 是一种热休克蛋白,已知其可使大的蛋白质复合物解聚。在集胞藻 PCC 6803 中组成型、16 倍过表达 ClpB1,当培养物快速(1°C/s)加热到 50°C 时,细胞存活率提高了 20 倍,在高温(>46°C)孵育时平均延迟细胞死亡 3 分钟。ClpB1 和另一种热休克蛋白 DnaK2 的共过表达进一步提高了细胞存活率。根据免疫细胞化学结果,ClpB1 分布在细胞质中,但在特定区域浓缩,并且在类囊体膜附近更为普遍。然而,ClpB1 的过表达不会导致形态、叶绿素含量或光系统比例发生变化。虽然电子显微镜显示在对照菌株中热处理后明显发生了蛋白质聚集,但在 ClpB1 过表达菌株中维持了蛋白质聚集体的大小。组成型 ClpB1 的过表达允许对热休克做出更早的反应,并防止培养物的快速加热。

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