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Bcl-2 Decreases the Affinity of SQSTM1/p62 to Poly-Ubiquitin Chains and Suppresses the Aggregation of Misfolded Protein in Neurodegenerative Disease

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Abstract

Poly-ubiquitinated protein aggregate formation is the most striking hallmark of various neurodegenerative diseases such as Alzheimer’s disease, Huntington’s disease, amyotrophic lateral sclerosis, and prion disease. Mutations of many ubiquitin-associated proteins involved in the regulation of protein aggregation, such as SQSTM1/p62 (p62), parkin, and VCP, are closely linked to neurodegeneration. B-cell lymphoma 2 (Bcl-2) is a key regulator in autophagy, apoptosis, and mitochondria quality control in many cell types including neurons, and it plays important roles in the pathogenesis of neurodegenerative diseases mentioned above. Our previous work showed that Bcl-2 can directly bind to p62, and here we report that Bcl-2 directly interacts with the N-terminus of p62, but not the C-terminus (UBA domain). Interestingly and importantly, Bcl-2 affects the affinity of p62 to poly-ubiquitin chains and suppresses the aggregation of poly-ubiquitinated proteins such as mutant huntingtin associated with Huntington’s disease. Our study reveals a role of Bcl-2 that involves in the regulation of misfolded proteins.

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Acknowledgments

This work was supported in part by the National High-tech Research and Development program of China 973-projects (2011CB504102), the National Natural Sciences Foundation of China (Nos. 31200803 and 31371072), Natural Science Foundation of Jiangsu Province (BK2012181), a project funded by Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases (BM2013003), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. Laboratory of Molecular Neuropathology, Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, 215021, China

    Liang Zhou, Hongfeng Wang, Haigang Ren, Qingsong Hu, Zheng Ying & Guanghui Wang

  2. Key Laboratory of Brain Function and Disease, School of Life Sciences, University of Science and Technology of China, Chinese Academy of Sciences, Hefei, Anhui, 230027, China

    Liang Zhou

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  1. Liang Zhou
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  3. Haigang Ren
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  4. Qingsong Hu
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Correspondence to Zheng Ying or Guanghui Wang.

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Liang Zhou and Hongfeng Wang contributed equally to this work.

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Fig. S1

H1299 cells were transfected with EGFP-Bcl-2 for 48 h, and then the cells were fixed and subjected to immunofluorescent assay using anti-p62 (Enzo life) antibody to detect endogenous p62 (red) and DAPI to stain the nuclei (blue). The scale bar indicates 10 μm. (DOCX 810 kb)

Fig. S2

HEK293 cells were transfected with EGFP-Htt-60Q and HA-ubiquitin (Ub) for 12 h, then the cells were subjected to immunofluorescent assay using anti-HA antibody (blue). The arrows indicate that Htt-60Q and ubiquitin could co-aggregate together. The scale bar indicates 10 μm. (DOCX 1387 kb)

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Zhou, L., Wang, H., Ren, H. et al. Bcl-2 Decreases the Affinity of SQSTM1/p62 to Poly-Ubiquitin Chains and Suppresses the Aggregation of Misfolded Protein in Neurodegenerative Disease. Mol Neurobiol 52, 1180–1189 (2015). https://doi.org/10.1007/s12035-014-8908-1

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