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BAG2 prevents Tau hyperphosphorylation and increases p62/SQSTM1 in cell models of neurodegeneration

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Abstract

Background

Protein aggregates are pathological hallmarks of many neurodegenerative diseases, however the physiopathological role of these aggregates is not fully understood. Protein quality control has a pivotal role for protein homeostasis and depends on specific chaperones. The co-chaperone BAG2 can target phosphorylated Tau for degradation by an ubiquitin-independent pathway, although its possible role in autophagy was not yet elucidated. In view of this, the aim of the present study was to investigate the association among protein aggregation, autophagy and BAG2 levels in cultured cells from hippocampus and locus coeruleus as well as in SH-SY5Y cell line upon different protein aggregation scenarios induced by rotenone, which is a flavonoid used as pesticide and triggers neurodegeneration.

Methods and results

The present study showed that rotenone exposure at 0.3 nM for 48 h impaired autophagy prior to Tau phosphorylation at Ser199/202 in hippocampus but not in locus coeruleus cells, suggesting that distinct neuron cells respond differently to rotenone toxicity. Rotenone induced Tau phosphorylation at Ser199/202, together with a decrease in the endogenous BAG2 protein levels in SH-SY5Y and hippocampus cell culture, which indicates that rotenone and Tau hyperphosphorylation can affect this co-chaperone. Finally, it has been shown that BAG2 overexpression, increased p62/SQSTM1 levels in cells from hippocampus and locus coeruleus, stimulated LC3II recycling as well as prevented the raise of phosphorylated Tau at Ser199/202 in hippocampus.

Conclusions

Results demonstrate a possible role for BAG2 in degradation pathways of specific substrates and its importance for the study of cellular aspects of neurodegenerative diseases.

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Data availability

Data will be available upon request.

Code availability

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Acknowledgements

Authors are grateful to Professor Alberto Ribeiro and Waldir Caldeira for their kind assistance in providing infrastructure and expertise to acquire confocal images. This study was supported by research grants from Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) (2011/06434-7; 2013/08028-1; 2018/07592-4). R.S.L. received fellowships from FAPESP (2016/04409-9 and 2017/24722-6).

Funding

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) (2011/06434-7; 2013/08028-1; 2018/07592-4; 2016/04409-9 and 2017/24722-6).

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Authors and Affiliations

  1. Departamento de Genetica e Biologia Evolutiva, Instituto de Biociencias, Universidade de Sao Paulo, Rua do Matao, 277, Cidade Universitaria, Sao Paulo, SP, 05508-090, Brazil

    Raquel S. Lima & Merari F. R. Ferrari

  2. Centro de Ciencias Naturais e Humanas, Universidade Federal do ABC, Santo Andre, SP, Brazil

    Daniel C. Carrettiero

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  1. Raquel S. Lima
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Contributions

RSL performed the experiments; RSL, DCC and MFRF analysed data and wrote the manuscript; MFRF supervised the study.

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Correspondence to Merari F. R. Ferrari.

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Authors declare that there are no conflicts of interest to be reported.

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All the procedures were performed in strict accordance with Institutional and International Guidelines for animal care and use [29], as well as respecting the Brazilian federal law 11794/08 for animal welfare and approved by the institutional ethics committee (CEUA 271/2016) of the Department of Genetics and Evolutionary Biology, Institute for Biosciences, University of Sao Paulo.

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11033_2022_7577_MOESM1_ESM.jpg

Supplementary Figure 1 (S1): Detailed protocol depicting the time course of culture, BAG-2 transfection, rotenone or chloroquine (CQ) exposure and analysis (JPG 643 kb)

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Lima, R.S., Carrettiero, D.C. & Ferrari, M.F.R. BAG2 prevents Tau hyperphosphorylation and increases p62/SQSTM1 in cell models of neurodegeneration. Mol Biol Rep 49, 7623–7635 (2022). https://doi.org/10.1007/s11033-022-07577-w

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