Skip to main content
SpringerLink
Log in

Molecular chaperones HSP40, HSP70, STIP1, and HSP90 are involved in stabilization of Cx43

  • Research
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

To investigate the involvement of stress induced phosphoprotein 1 (STIP1), heat shock protein (HSP) 70, and HSP90 in ubiquitination of connexin 43 (Cx43) in rat H9c2 cardiomyocytes. Co-immunoprecipitation was used to detect protein-protein interactions and Cx43 ubiquitination. Immunofluorescence was used for protein co-localization. The protein binding, Cx43 protein expression, and Cx43 ubiquitination were reanalyzed in H9c2 cells with modified STIP1 and/or HSP90 expression. STIP1 bound to HSP70 and HSP90, and Cx43 bound to HSP40, HSP70, and HSP90 in normal H9c2 cardiomyocytes. Overexpression of STIP1 promoted the transition of Cx43-HSP70 to Cx43-HSP90 and inhibited Cx43 ubiquitination; knockdown of STIP1 resulted in the opposite effects. Inhibition of HSP90 counteracted the inhibitory effect of STIP1 overexpression on Cx43 ubiquitination. STIP1 suppresses Cx43 ubiquitination in H9c2 cardiomyocytes by promoting the transition of Cx43-HSP70 to Cx43-HSP90.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

References

  • Altinok S, Sanchez-Hodge R, Stewart M, Smith K, Schisler JC (2021) With or without you: co-chaperones mediate health and disease by modifying chaperone function and protein triage. Cells 10:3121

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Baker HA, Bernardini JP (2021) It’s not just a phase; ubiquitination in cytosolic protein quality control. Biochem Soc Trans 49:365–377

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Boengler K, Rohrbach S, Weissmann N, Schulz R (2021) Importance of Cx43 for right ventricular function. Int J Mol Sci 22:987

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brundel B (2020) The role of proteostasis derailment in cardiac diseases. Cells 9:2317

    Article  PubMed  PubMed Central  Google Scholar 

  • Carroll EC, Marqusee S (2022) Site-specific ubiquitination: deconstructing the degradation tag. Curr Opin Struct Biol 73:102345

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen B, Feder ME, Kang L (2018) Evolution of heat-shock protein expression underlying adaptive responses to environmental stress. Mol Ecol 27:3040–3054

    Article  PubMed  Google Scholar 

  • da Fonseca ACC, Matias D, Geraldo LHM, Leser FS, Pagnoncelli I, Garcia C, do Amaral RF, da Rosa BG, Grimaldi I, de Magalhaes ES, Coppola-Segovia V, de Azevedo EM, Zanata SM, Lima FRS (2021) The multiple functions of the co-chaperone stress inducible protein 1. Cytokine Growth Factor Rev 57:73–84

    Article  PubMed  Google Scholar 

  • Dietrich M, Malik MS, Nikolaysen F, Skeie M, Stang E (2018) Protein kinase C mediated internalization of ErbB2 is independent of clathrin, ubiquitination and Hsp90 dissociation. Exp Cell Res 371:139–150

    Article  CAS  PubMed  Google Scholar 

  • Epifantseva I, Shaw RM (2018) Intracellular trafficking pathways of Cx43 gap junction channels. Biochim Biophys Acta Biomembr 1860:40–47

    Article  CAS  PubMed  Google Scholar 

  • Faust O, Abayev-Avraham M, Wentink AS, Maurer M, Nillegoda NB, London N, Bukau B, Rosenzweig R (2020) HSP40 proteins use class-specific regulation to drive HSP70 functional diversity. Nature 587:489–494

    Article  CAS  PubMed  Google Scholar 

  • Genest O, Wickner S, Doyle SM (2019) Hsp90 and Hsp70 chaperones: collaborators in protein remodeling. J Biol Chem 294:2109–2120

    Article  CAS  PubMed  Google Scholar 

  • Ghosh A, Dai Y, Biswas P, Stuehr DJ (2019) Myoglobin maturation is driven by the hsp90 chaperone machinery and by soluble guanylyl cyclase. FASEB J 33:9885–9896

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hernandez MP, Sullivan WP, Toft DO (2002) The assembly and intermolecular properties of the hsp70-Hop-hsp90 molecular chaperone complex. J Biol Chem 277:38294–38304

    Article  CAS  PubMed  Google Scholar 

  • Himelman E, Lillo MA, Nouet J, Gonzalez JP, Zhao Q, Xie LH, Li H, Liu T, Wehrens XH, Lampe PD, Fishman GI, Shirokova N, Contreras JE, Fraidenraich D (2020) Prevention of connexin-43 remodeling protects against Duchenne muscular dystrophy cardiomyopathy. J Clin Invest 130:1713–1727

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jang J, Lee SH, Kang DH, Sim DW, Ryu KS, Jo KS, Lee J, Ryu H, Kim EH, Won HS, Kim JH (2022) Structural resemblance of the DNAJA-family protein, Tid1, to the DNAJB-family Hsp40. BMB Rep 55:488–493

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lackie RE, Maciejewski A, Ostapchenko VG, Marques-Lopes J, Choy WY, Duennwald ML, Prado VF, Prado MAM (2017) The Hsp70/Hsp90 chaperone machinery in neurodegenerative diseases. Front Neurosci 11:254

    Article  PubMed  PubMed Central  Google Scholar 

  • Lackie RE, Razzaq AR, Farhan SMK, Qiu LR, Moshitzky G, Beraldo FH, Lopes MH, Maciejewski A, Gros R, Fan J, Choy WY, Greenberg DS, Martins VR, Duennwald ML, Lerch JP, Soreq H, Prado VF, Prado MAM (2020) Modulation of hippocampal neuronal resilience during aging by the Hsp70/Hsp90 co-chaperone STI1. J Neurochem 153:727–758

    Article  CAS  PubMed  Google Scholar 

  • Leutert M, Entwisle SW, Villen J (2021) Decoding post-translational modification crosstalk with proteomics. Mol Cell Proteomics 20:100129

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li Y, Li S, Wu H (2022) Ubiquitination-proteasome system (UPS) and autophagy two main protein degradation machineries in response to cell stress. Cells 11:851

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu Q, Liang C, Zhou L (2020) Structural and functional analysis of the Hsp70/Hsp40 chaperone system. Protein Sci 29:378–390

    Article  CAS  PubMed  Google Scholar 

  • Martins-Marques T, Catarino S, Goncalves A, Miranda-Silva D, Goncalves L, Antunes P, Coutinho G, Leite Moreira A, Falcao I, Pires, Girao H (2020) EHD1 modulates Cx43 gap junction remodeling associated with cardiac diseases. Circ Res 126:e97–e113

    Article  CAS  PubMed  Google Scholar 

  • Moran Luengo T, Mayer MP, Rudiger SGD (2019) The Hsp70-Hsp90 chaperone cascade in protein folding. Trends Cell Biol 29:164–177

    Article  CAS  PubMed  Google Scholar 

  • Pecoraro M, Pala B, Di Marcantonio MC, Muraro R, Marzocco S, Pinto A, Mincione G, Popolo A (2020) Doxorubicininduced oxidative and nitrosative stress: mitochondrial connexin 43 is at the crossroads. Int J Mol Med 46:1197–1209

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pernot M, Jaspard-Vinassa B, Abelanet A, Rubin S, Forfar I, Jeanningros S, Cetran L, Yu MH, Balse E, Hatem S, Dufourcq P, Couffinhal T, Duplaa C (2022) Decrease of Pdzrn3 is required for heart maturation and protects against heart failure. Sci Rep 12:8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rape M (2018) Ubiquitylation at the crossroads of development and disease. Nat Rev Mol Cell Biol 19:59–70

    Article  CAS  PubMed  Google Scholar 

  • Rodriguez-Sinovas A, Boengler K, Cabestrero A, Gres P, Morente M, Ruiz-Meana M, Konietzka I, Miro E, Totzeck A, Heusch G, Schulz R, Garcia-Dorado D (2006) Translocation of connexin 43 to the inner mitochondrial membrane of cardiomyocytes through the heat shock protein 90-dependent TOM pathway and its importance for cardioprotection. Circ Res 99:93–101

    Article  CAS  PubMed  Google Scholar 

  • Rohl A, Wengler D, Madl T, Lagleder S, Tippel F, Herrmann M, Hendrix J, Richter K, Hack G, Schmid AB, Kessler H, Lamb DC, Buchner J (2015) Hsp90 regulates the dynamics of its cochaperone Sti1 and the transfer of Hsp70 between modules. Nat Commun 6:6655

    Article  PubMed  Google Scholar 

  • Rosenzweig R, Nillegoda NB, Mayer MP, Bukau B (2019) The Hsp70 chaperone network. Nat Rev Mol Cell Biol 20:665–680

    Article  CAS  PubMed  Google Scholar 

  • Ruckova E, Muller P, Nenutil R, Vojtesek B (2012) Alterations of the Hsp70/Hsp90 chaperone and the HOP/CHIP co-chaperone system in cancer. Cell Mol Biol Lett 17:446–458

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Soejima M, Koda Y (2008) TaqMan-based real-time PCR for genotyping common polymorphisms of haptoglobin (HP1 and HP2). Clin Chem 54:1908–1913

    Article  CAS  PubMed  Google Scholar 

  • Tang J, Yan T, Bao Y, Shen C, Yu C, Zhu X, Tian X, Guo F, Liang Q, Liu Q, Zhong M, Chen J, Ge Z, Li X, Chen X, Cui Y, Chen Y, Zou W, Chen H, Hong J, Fang JY (2019) LncRNA GLCC1 promotes colorectal carcinogenesis and glucose metabolism by stabilizing c-Myc. Nat Commun 10:3499

    Article  PubMed  PubMed Central  Google Scholar 

  • Totland MZ, Rasmussen NL, Knudsen LM, Leithe E (2020) Regulation of gap junction intercellular communication by connexin ubiquitination: physiological and pathophysiological implications. Cell Mol Life Sci 77:573–591

    Article  CAS  PubMed  Google Scholar 

  • Wang JD, Shao Y, Liu D, Liu NY, Zhu DY (2021a) Rictor/mTORC2 involves mitochondrial function in ES cells derived cardiomyocytes via mitochondrial connexin 43. Acta Pharmacol Sin 42:1790–1797

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Y, Wang Y, Li F, Zhang X, Li H, Yang G, Xu C, Wei C (2021) Spermine protects cardiomyocytes from high glucose-induced energy disturbance by targeting the CaSR-gp78-ubiquitin proteasome system. Cardiovasc Drugs Ther 35:73–85

    Article  CAS  PubMed  Google Scholar 

  • Xie J, Li H, Li S, Li J, Li Y (2022) Molecular mechanism of sevoflurane preconditioning based on whole-transcriptome sequencing of lipopolysaccharide-induced cardiac dysfunction in mice. J Cardiovasc Pharmacol 79:846–857

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Thanks for all the contributors.

Funding

This study were funded by the National Natural Science Foundation (NSFC) Regional Fund Cultivation Program of Affiliated Hospital of Guizhou Medical University (No. gyfynsfc[2022]-47); the Science and Technology Fund Project of Guizhou Provincial Health and Health Commission (No. gzwkj2022-120).

Author information

Authors and Affiliations

  1. Guizhou Medical University, Guiyang, 550004, Guizhou, People’s Republic of China

    Li An

  2. Department of Anesthesiology, The Affiliated Hospital of Guizhou Medical University, No. 28 Guiyi St, Yunyan District, Guiyang, 550004, Guizhou, People’s Republic of China

    Li An, Hong Gao, Yi Zhong & Jing Yi

  3. Translational Medicine Research Center of Guizhou Medical University, Guiyang, 550004, Guizhou, People’s Republic of China

    Li An

  4. Department of Anesthesiology, Guiyang Fourth People’s Hospital, Guiyang, 550002, Guizhou, People’s Republic of China

    Yanqiu Liu

  5. School of Anesthesiology, Guizhou Medical University, Guiyang, 550004, Guizhou, People’s Republic of China

    Ying Cao, Xiang Huang, Rui Tong, Zhijun Pan, Xu Yan, Meiyan Liu, Shengzhao Wang, Xue Bai, Hao Wu & Tingju Hu

  6. Guiyang Second People’s Hospital, Guiyang, 550001, Guizhou, People’s Republic of China

    Ying Cao

  7. Children’s Hospital of Guiyang Maternal and Child Health Hospital, Guiyang, 550001, Guizhou, People’s Republic of China

    Chunlei Wen

Authors
  1. Li An
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hong Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yi Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yanqiu Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ying Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jing Yi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xiang Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Chunlei Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Rui Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Zhijun Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Xu Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Meiyan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Shengzhao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Xue Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Hao Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Tingju Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

GH and AL conceived the ideas. GH and AL designed the experiments. AL; ZY; LYQ; CY and YJ performed the experiments. AL; ZY; HX; WCL; TR; PZJ; YX; LMY; WSZ; BX; WH and HTJ analyzed the data. AL; LYQ and CY provided critical materials. TR; PZJ; YX and LMY wrote the manuscript. GH supervised the study. All the authors have read and approved the final version for publication.

Corresponding author

Correspondence to Hong Gao.

Ethics declarations

Competing interests

The authors declare there is no conflict of interests.

Ethical approval

This research didn’t contain the experiments involved in animals or human participants, there is no need for ethical statement.

Consent for publication

Not applicable.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

An, L., Gao, H., Zhong, Y. et al. Molecular chaperones HSP40, HSP70, STIP1, and HSP90 are involved in stabilization of Cx43. Cytotechnology 75, 207–217 (2023). https://doi.org/10.1007/s10616-023-00570-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-023-00570-6

Keywords

Search

Navigation