Heat shock proteins and protein quality control: implications for protein misfolding diseases and cellular and organismal ageing
All eukaryotic cells contain evolutionarily highly conserved systems to combat proteotoxic stress one of which is the heat shock (HS). The integrity of the HS system is crucial for cell survival because they ensure the quality control of cellular proteins with respect to structure as well as synthesis and degradation (so-called protein homeostasis). Failure of the quality control mechanism is involved in many age-related syndromes, including neurodegenerative diseases. By boosting the stress system with age, one might be to dampen the adverse effect of genetic mutations and destabilizing protein modifications and thus ameliorate or prevent age-related disease. Practical application is, however, still hampered by lack of knowledge of the stress system and its precise function and regulation within the living cell.
- To understand the functioning and regulation (by cofactors) of the diverse chaperone machines in living mammalian cells and to know the nature and extent of redundancy in the total chaperone network of a cell to be able to determine the optimal target for intervention in the stress system.
- To understand the (putative) roles that heat shock proteins may have in human diseases, in particular neurodegenerative diseases such as Huntington's disease, and ageing in general.
- Role of chaperones in the proteasomal and autophagosomal protein degradation routes
- Regulation of storage of misfolded proteins and their handling by chaperones
- Role of DNAJB6 in amelioration of polyglutamine diseases
- Role of the HSPB8/BAG3 complex in astrocyte-neuronal interactions in disease
- Screens for protective chaperones in Parkinson disease and Amyotrofic laterale sclerosis
- Compound screens for modifyiers of chaperone activity
- Chaperones and longevity
Techniques and approaches
The above-mentioned goals are mainly approached by cellular model systems in which we try to manipulated Hsp-expressions by regulated overexpression and RNA interference. The work includes molecular cloning, gene transfections, reporter systems (luminicence), a variety of molecular/biochemical assays (SDS-PAGE, Westerns, and Notherns), cellular chaperone measurements and a variety of molecular cell biological techniques (immunohistochemistry, confocal microscopy, live-recording using GFP-tagged proteins). Genomic and proteomics experiments are being planned for to investigate the nature and extent of redundancy in the total chaperone network of a cell, and hence determine the optimal target for intervention in the stress system. Finally, Drosophila melanogaster is used as a simple in vivo model for protein folding diseases, chaperonopathies (i.e. disease related to mutations in Hsp), and ageing.
Carra S, Crippa V,Rusmini P, Boncoraglio A, Minoia M, Giorgetti E; Kampinga HH; Poletti A. Alteration of protein folding and degradation in motor neuron diseases: implications and protective functions of Small Heat Shock proteins. Progress in Neurobiology (2012) in press
Vos MJ, Zijlstra MP, Carra S, Sibon OCM, Kampinga HH. Small heat shock proteins, protein degradation and protein aggregation diseases. Autophagy 7 (2011) 101-103.*
Kampinga HH, Craig EA. The HSP70 chaperone machinery: J proteins as drivers of functional specificity. Nature Reviews Mol. Cell. Biol. 11 (2010) 579-592.
Hageman J, Rujano MA, van Waarde MAWH, Kakkar V, Dirks D, Govorukhina N, Oosterveld-Hut HJM, Lubsen NH, Kampinga HH. A DNAJB Chaperone Subfamily with HDAC-dependent Activities Suppresses Toxic Protein Aggregation. Molecular Cell 37 (2010) 355-369.
Vos MJ, Zijlstra MP, Kanon B, van Waarde-Verhagen MAWH, Brunt ERP, Oosterveld-Hut HMJ, Carra S, Sibon OCM, Kampinga HH. HSPB7 is the most potent polyQ aggregation suppressor within the HSPB family of molecular chaperones. Human Molecular Genetics 19 (2010) 4677-4693.
Carra S, Boncoraglio A, Bart Kanon, Brunsting JF, Minoia M, Rana A, Vos MJ, Seidel K, Sibon OCM, Kampinga HH. Identification of the Drosophila ortholog of HSPB8: implication of HSPB8 loss of function in protein folding diseases. J Biol Chem. 285 (2010) 37811-37822.
Zijlstra MP, Rujano MA, van Waarde MAWH, Vis E, Brunt ERP,Kampinga HH., Levels of DNAJB family members (HSP40) correlate with disease onset in patients with spinocerebellar ataxia type 3. Eur. J. Neuroscience 32 (2010) 760–770.
Carra S, Brunsting JF, Lambert H, Landry J, Kampinga HH, HSPB8 participates in protein quality control by a non chaperone-like mechanism that requires eIF2alpha phosphorylation. J Biol Chem. 284 (2009) 5523-5532.
Hageman J and Kampinga HH, Computational analysis of the human HspH/HspA/DnaJ family and Cloning of a human HspH/HspA/DnaJ expression library. Cell Stress & Chaperones. 14 (2009) 1-21.
Kampinga HH, Hageman J, Vos MJ, Kubota H, Tanguay RM, Bruford EA, Cheetham ME, Chen B, Hightower LE. Guidelines for the nomenclature of the human heat shock proteins. Cell Stress & Chaperones. 14 (2009) 105-111.
Vos MJ, Hageman J, Carra S, and Kampinga HH, Structural and functional diversities between members of the human HspB, HspH, HspA and DnaJ chaperone families. Biochemistry 47 (2008) 7001-7011.
- Senter Novem (IOP genomics) grant (project nr. IGE07004)
- Hersenstichting Nederland
- National Ataxia Foundation
- Prinses Beatrix Foundation
- AFM trampoline Grant SNN project (Transitie II & Pieken)
|Last modified:||January 24, 2015 11:59|