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Osmotic stress-induced, rapid clustering of IGF2BP proteins nucleates stress granule assembly

View ORCID ProfileWei-jie Zeng, Chuxin Lu, Yuanyuan Shi, Xinxin Chen, View ORCID ProfileJie Yao
doi: https://doi.org/10.1101/846659
Wei-jie Zeng
1Sun Yat-sen University School of Medicine, Guangzhou, 510275, China
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Chuxin Lu
1Sun Yat-sen University School of Medicine, Guangzhou, 510275, China
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Yuanyuan Shi
1Sun Yat-sen University School of Medicine, Guangzhou, 510275, China
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Xinxin Chen
1Sun Yat-sen University School of Medicine, Guangzhou, 510275, China
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Jie Yao
1Sun Yat-sen University School of Medicine, Guangzhou, 510275, China
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  • For correspondence: jie.yao@alleninstitute.org
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Abstract

Stress granules (SGs) are formed in the cytoplasm by liquid-liquid phase separation (LLPS) of translationally-stalled mRNA and RNA-binding proteins during stress response. Understanding the mechanisms governing SG assembly requires imaging SG formation in real time. Here we used live cell imaging and super-resolution imaging to visualize SG assembly in human cells. We found that IGF2BP proteins formed microscopically visible clusters almost instantaneously upon osmotic stress, prior to the recruitment of G3BP1 and TIA1. The rapid clustering of IGF2BP1 was ATP-independent and was mediated by its KH3/4 di-domains and an intrinsically disordered region (IDR), whereas ATP depletion inhibited the recruitment of G3BP1 and TIA1. Moreover, we detected cytoplasmic clusters of IGF2BP1 below the optical resolution in normal cells and found IGF2BP1 forming a dense granule associated with multiple clusters of poly(A) mRNA in mature SGs. Thus, ATP-independent, rapid clustering of IGF2BP nucleates SG assembly during osmotic stress.

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  • 20191202 revised - complete version

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Posted December 02, 2019.
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Osmotic stress-induced, rapid clustering of IGF2BP proteins nucleates stress granule assembly
Wei-jie Zeng, Chuxin Lu, Yuanyuan Shi, Xinxin Chen, Jie Yao
bioRxiv 846659; doi: https://doi.org/10.1101/846659
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Osmotic stress-induced, rapid clustering of IGF2BP proteins nucleates stress granule assembly
Wei-jie Zeng, Chuxin Lu, Yuanyuan Shi, Xinxin Chen, Jie Yao
bioRxiv 846659; doi: https://doi.org/10.1101/846659

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