Cell Stress and Chaperones

, Volume 22, Issue 4, pp 531–540 | Cite as

An interaction study in mammalian cells demonstrates weak binding of HSPB2 to BAG3, which is regulated by HSPB3 and abrogated by HSPB8

  • Federica F. Morelli
  • Laura Mediani
  • Lonneke Heldens
  • Jessika Bertacchini
  • Ilaria Bigi
  • Arianna Dorotea Carrà
  • Jonathan Vinet
  • Serena Carra
SMALL HEAT SHOCK PROTEINS

Abstract

The ten mammalian small heat shock proteins (sHSPs/HSPBs) show a different expression profile, although the majority of them are abundant in skeletal and cardiac muscles. HSPBs form hetero-oligomers and homo-oligomers by interacting together and complexes containing, e.g., HSPB2/HSPB3 or HSPB1/HSPB5 have been documented in mammalian cells and muscles. Moreover, HSPB8 associates with the Hsc70/Hsp70 co-chaperone BAG3, in mammalian, skeletal, and cardiac muscle cells. Interaction of HSPB8 with BAG3 regulates its stability and function. Weak association of HSPB5 and HSPB6 with BAG3 has been also reported upon overexpression in cells, supporting the idea that BAG3 might indirectly modulate the function of several HSPBs. However, it is yet unknown whether other HSPBs highly expressed in muscles such as HSPB2 and HSPB3 also bind to BAG3. Here, we report that in mammalian cells, upon overexpression, HSPB2 binds to BAG3 with an affinity weaker than HSPB8. HSPB2 competes with HSPB8 for binding to BAG3. In contrast, HSPB3 negatively regulates HSPB2 association with BAG3. In human myoblasts that express HSPB2, HSPB3, HSPB8, and BAG3, the latter interacts selectively with HSPB8. Combining these data, it supports the interpretation that HSPB8-BAG3 is the preferred interaction.

Keywords

Small heat shock proteins/HSPBs BAG3 Interaction Competition 

Notes

Acknowledgements

We thank Prof. E. Pegoraro and Dr. E. Galletta for providing the LHCNM2 cells and for useful technical support for their use. SC is grateful to Prof. Harm H. Kampinga for useful discussions and tools. SC is grateful to Telethon (GEP12008 and GGP15001) and Association Francaise contres les Myopathies (grant number 15999) for financial support. SC also thanks the Centro Interdipartimentale Grandi Strumenti (CIGS) of the University of Modena and Reggio Emilia for support with confocal microscopy.

References

  1. Arimura T, Ishikawa T et al (2011) Dilated cardiomyopathy-associated BAG3 mutations impair Z-disc assembly and enhance sensitivity to apoptosis in cardiomyocytes. Hum Mutat 32(12):1481–1491CrossRefPubMedGoogle Scholar
  2. Brady JP, Garland DL et al (2001) AlphaB-crystallin in lens development and muscle integrity: a gene knockout approach. Invest Ophthalmol Vis Sci 42(12):2924–2934PubMedGoogle Scholar
  3. Carra S, Boncoraglio A et al (2010) Identification of the drosophila ortholog of HSPB8: implication of HSPB8 loss of function in protein folding diseases. J Biol Chem 285(48):37811–37822CrossRefPubMedPubMedCentralGoogle Scholar
  4. Carra S, Seguin SJ et al (2008) HspB8 chaperone activity toward poly (Q)-containing proteins depends on its association with Bag3, a stimulator of macroautophagy. J Biol Chem 283(3):1437–1444CrossRefPubMedGoogle Scholar
  5. Chami N, Tadros R et al (2014) Nonsense mutations in BAG3 are associated with early-onset dilated cardiomyopathy in French Canadians. Can J Cardiol 30(12):1655–1661CrossRefPubMedGoogle Scholar
  6. den Engelsman J, Boros S et al (2009) The small heat-shock proteins HSPB2 and HSPB3 form well-defined heterooligomers in a unique 3 to 1 subunit ratio. J Mol Biol 393(5):1022–1032CrossRefGoogle Scholar
  7. Evgrafov OV, Mersiyanova I et al (2004) Mutant small heat-shock protein 27 causes axonal Charcot-Marie-tooth disease and distal hereditary motor neuropathy. Nat Genet 36(6):602–606CrossRefPubMedGoogle Scholar
  8. Fontaine JM, Sun X et al (2005) Interactions of HSP22 (HSPB8) with HSP20, alphaB-crystallin, and HSPB3. Biochem Biophys Res Commun 337(3):1006–1011CrossRefPubMedGoogle Scholar
  9. Franaszczyk M, Bilinska ZT et al (2014) The BAG3 gene variants in polish patients with dilated cardiomyopathy: four novel mutations and a genotype-phenotype correlation. J Transl Med 12:192CrossRefPubMedPubMedCentralGoogle Scholar
  10. Fuchs M, Poirier DJ et al (2010) Identification of the key structural motifs involved in HspB8/HspB6-Bag3 interaction. Biochem J 425(1):245–255CrossRefGoogle Scholar
  11. Ghaoui R, Palmio J et al (2015) Mutations in HSPB8 causing a new phenotype of distal myopathy and motor neuropathy. Neurology 86(4):391–398CrossRefPubMedGoogle Scholar
  12. Hishiya A, Salman MN et al (2011) BAG3 directly interacts with mutated alphaB-crystallin to suppress its aggregation and toxicity. PLoS One 6(3):e16828CrossRefPubMedPubMedCentralGoogle Scholar
  13. Irobi J, Van Impe K et al (2004) Hot-spot residue in small heat-shock protein 22 causes distal motor neuropathy. Nat Genet 36(6):597–601CrossRefPubMedGoogle Scholar
  14. Kolb SJ, Snyder PJ et al (2010) Mutant small heat shock protein B3 causes motor neuropathy: utility of a candidate gene approach. Neurology 74(6):502–506CrossRefPubMedGoogle Scholar
  15. Lam WY, Wing Tsui SK et al (1996) Isolation and characterization of a human heart cDNA encoding a new member of the small heat shock protein family—HSPL27. Biochim Biophys Acta 1314(1–2):120–124CrossRefPubMedGoogle Scholar
  16. Liu C, Welsh MJ (1999) Identification of a site of Hsp27 binding with Hsp27 and alpha B-crystallin as indicated by the yeast two-hybrid system. Biochem Biophys Res Commun 255(2):256–261CrossRefPubMedGoogle Scholar
  17. Norton N, Li D et al (2011) Genome-wide studies of copy number variation and exome sequencing identify rare variants in BAG3 as a cause of dilated cardiomyopathy. Am J Hum Genet 88(3):273–282CrossRefPubMedPubMedCentralGoogle Scholar
  18. Oshita SE, Chen F et al (2010) The small heat shock protein HspB2 is a novel anti-apoptotic protein that inhibits apical caspase activation in the extrinsic apoptotic pathway. Breast Cancer Res Treat 124(2):307–315CrossRefPubMedPubMedCentralGoogle Scholar
  19. Pennings JL, van Dartel DA et al (2011) Identification by gene coregulation mapping of novel genes involved in embryonic stem cell differentiation. Stem Cells Dev 20(1):115–126CrossRefPubMedGoogle Scholar
  20. Rauch, J. N., E. Tse, et al. (2016). BAG3 Is a Modular, Scaffolding Protein that physically Links Heat Shock Protein 70 (Hsp70) to the Small Heat Shock Proteins. J Mol Biol.Google Scholar
  21. Selcen D, Muntoni F et al (2009) Mutation in BAG3 causes severe dominant childhood muscular dystrophy. Ann Neurol 65(1):83–89CrossRefPubMedPubMedCentralGoogle Scholar
  22. Shemetov AA, Gusev NB (2011) Biochemical characterization of small heat shock protein HspB8 (Hsp22)-Bag3 interaction. Arch Biochem Biophys 513(1):1–9CrossRefPubMedGoogle Scholar
  23. Sugiyama Y, Suzuki A et al (2000) Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation. J Biol Chem 275(2):1095–1104CrossRefPubMedGoogle Scholar
  24. Sun X, Fontaine JM et al (2004) Interaction of human HSP22 (HSPB8) with other small heat shock proteins. J Biol Chem 279(4):2394–2402CrossRefPubMedGoogle Scholar
  25. Taipale M, Tucker G et al (2014) A quantitative chaperone interaction network reveals the architecture of cellular protein homeostasis pathways. Cell 158(2):434–448CrossRefPubMedPubMedCentralGoogle Scholar
  26. Toro R, Perez-Serra A et al (2016) Familial dilated cardiomyopathy caused by a novel Frameshift in the BAG3 Gene. PLoS One 11(7):e0158730CrossRefPubMedPubMedCentralGoogle Scholar
  27. Vicart P, Caron A et al (1998) A missense mutation in the alphaB-crystallin chaperone gene causes a desmin-related myopathy. Nat Genet 20(1):92–95CrossRefPubMedGoogle Scholar
  28. Villard E, Perret C et al (2011) A genome-wide association study identifies two loci associated with heart failure due to dilated cardiomyopathy. Eur Heart J 32(9):1065–1076CrossRefPubMedPubMedCentralGoogle Scholar
  29. Vos MJ, Zijlstra MP et al (2010) HSPB7 is the most potent polyQ aggregation suppressor within the HSPB family of molecular chaperones. Hum Mol Genet 19(23):4677–4693CrossRefPubMedGoogle Scholar
  30. Zantema A, Verlaan-De Vries M et al (1992) Heat shock protein 27 and alpha B-crystallin can form a complex, which dissociates by heat shock. J Biol Chem 267(18):12936–12941PubMedGoogle Scholar

Copyright information

© Cell Stress Society International 2017

Authors and Affiliations

  1. 1.Centre for Neuroscience and Nanotechnology, Department of Biomedical, Metabolic and Neural SciencesUniversity of Modena and Reggio EmiliaModenaItaly
  2. 2.Department of Cell BiologyUniversity Medical Center Groningen; University of GroningenGroningenThe Netherlands

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