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Heat shock protein 70 inhibits hydrogen peroxide-induced nucleolar fragmentation via suppressing cleavage and down-regulation of nucleolin

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Cell Stress and Chaperones Aims and scope

Abstract

It has been reported that nucleolar fragmentation is a part of the overall apoptotic morphology, however, it is currently obscure whether and how nucleolar fragmentation can be induced by hydrogen peroxide (H2O2) and heat shock protein 70 (Hsp70) can prevent nucleolar fragmentation. To dissect these two questions, C2C12 myogenic cells and immortalized mouse embryonic fibroblasts (MEFs) with heat shock transcriptional factor 1 (HSF1) null mutation were treated with heat shock response (HS) (42.5 ± 0.5°C for 1 h and recovery at 37°C for 24 h) and then were insulted with 0.5 mmol/L H2O2. Morphological changes of nucleoli were observed under contrast microscope or electronic microscope. It was found that (1) stimulation with H2O2-induced nucleolar fragmentation by mediating cleavage and down-regulation of nucleolar protein, nucleolin in C2C12 myocytes and MEFs; (2) HS suppressed nucleolar fragmentation by inducing the expression of Hsp70 in an HSF1-dependent manner as indicated by assays of transfection with Hsp70 antisense oligonucleotides (AS-ONs) or recombinant plasmids of full-length Hsp70 cDNA; (3) protection of Hsp70 against nucleolar fragmentation was related to its accumulation in nucleolus mediated by nuclear localization sequence and its inhibition against cleavage and down-regulation of nucleolin. These results suggested that H2O2-induced nucleolar fragmentation and HS or Hsp70 inhibit H2O2-induced nucleolar fragmentation through the translocation of Hsp70 into nucleolar and its protection against impairment of nucleolin.

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Abbreviations

AS-ONs:

Antisense oligonucleotides

S-ONs:

Sense oligonucleotides

HS:

Heat shock response

HSF1:

Heat shock transcriptional factor 1

Hsps:

Heat shock proteins

Hsp70:

Heat shock protein 70

H2O2 :

Hydrogen peroxide

MEFs:

Mouse embryonic fibroblasts

NLS:

Nuclear localization sequence

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Acknowledgment

We acknowledge the kind gifts of HSF1 knock-out mice and pBR-HSP70 plasmid from Dr. Ivor Benjamin. We thank for Dr. McMillan DR critically reading the manuscript. This work was supported by grants from the Major National Basic Research Program of China (2007CB512007) and Natural Science Foundation of Hunan Province (11JJ2047).

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Authors and Affiliations

  1. Department of Pathophysiology, Xiangya School of Medicine, Central South University, 110 Xiangya Road, Changsha, Hunan, 410008, People’s Republic of China

    Kangkai Wang, Gonghua Deng, Guangwen Chen, Meidong Liu, Yuxin Yi & Xiangzhong Xiao

  2. School of Public Health, Central South University, Changsha, Hunan, 410008, People’s Republic of China

    Tubao Yang

  3. Division of Pediatric Endocrinology, Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX, 75390-9063, USA

    Daniel R. McMillan

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  1. Kangkai Wang
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  2. Gonghua Deng
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Correspondence to Xiangzhong Xiao.

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Wang, K., Deng, G., Chen, G. et al. Heat shock protein 70 inhibits hydrogen peroxide-induced nucleolar fragmentation via suppressing cleavage and down-regulation of nucleolin. Cell Stress and Chaperones 17, 121–130 (2012). https://doi.org/10.1007/s12192-011-0292-4

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