Rôle des protéines de choc thermique HSP90 et HSP70 dans la différenciation macrophagique

Abstract : Heat shock proteins (HSPs) are molecular chaperones whose expression is increased after many different stresses. They have a protective function helping the cell to cope with lethal conditions. These proteins play an essential role as molecular chaperones by assisting the correct folding of nascent and stress-accumulated misfolded proteins and by preventing their aggregation. My team is interested in understanding the roles of HSPs in two physiological related processes: apoptosis and cell differentiation. The aim of my work is to study the functions of HSP90 and HSP70 in macrophagic differentiation. I first studied the role of HSP90 in macrophagic differentiation. We previously reported that cellular inhibitor of apoptosis protein-1 (c-IAP1), migrated from the nucleus to the surface of the Golgi apparatus in cells undergoing differentiation. c-IAP1 is a member of the inhibitor of apoptosis protein (IAP) family which has demonstrated functions in cell death, cell signaling and mitosis. Here, we show that c-IAP1 is a client protein of the stress protein HSP90β. In three distinct cellular models, the two proteins interact and migrate from the nucleus to the cytoplasm during the differentiation process through a leptomycin B-sensitive pathway. Inhibition of HSP90 proteins by small chemical molecules and specific depletion of HSP90isoform by siRNA both leads to c-IAP1 degradation by the proteasomal machinery. This chaperone function of HSP90 towards c-IAP1 is specific of its β isoform as specific depletion of HSP90α isoform does not affect c-IAP1 content. Chemical inhibition of HSP90 or siRNA-mediated depletion of HSP90both inhibit cell differentiation, which can be reproduced by siRNA-mediated depletion of c-IAP1. Altogether, these results suggest that HSP90prevents auto-ubiquitination and degradation of its client protein c-IAP1, whose depletion would be sufficient to inhibit cell differentiation. The second part of my work consisted in the study of the role of HSP70 in macrophagic differentiation. We used two models of differentiation: human peripheral monocytes exposed to M-CSF and THP1 cells induced to differentiate by TPA. We found that in both models, HSP70 expression was induced during differentiation. Interestingly, the expression of Spi-1/Pu.1, a transcription factor essential for monocytes to differentiate, was similarly induced. Upon differentiation, both proteins co-localized in the nucleus and associated. Inhibition or down regulation of HSP70 induced Spi-1/Pu.1 degradation and blocks the differentiation process, indicating that the necessity of Spi-1/Pu.1 to be chaperoned by HSP70 during differentiation. Since Spi-1/Pu.1 promoter has a HSE-like, we studied whether transcription factors responsible for HSPs induction, HSF, could be involved. We show that although HSF2 do not seem involved, HSF1 binds to Spi-1/Pu.1 promoter and its inhibition blocks Spi-1/Pu.1 expression and monocytes differentiation. So HSP90 and HSP70 are essentials for macrophagic differentiation. Understanding monocyte differentiation regulation is important since defects of the differentiation process can lead to the development of leukemias (acute myeloid leukemia, chronic myelomonocytic leukemia). A better understanding of the roles of HSPs may provide new therapeutic strategies for the treatment of these pathologies.
Document type :
Sciences du Vivant [q-bio]. Université de Bourgogne, 2010. Français. <NNT : 2010DIJOS007>

Contributor : Abes Star <>
Submitted on : Friday, January 28, 2011 - 3:39:06 PM
Last modification on : Saturday, September 6, 2014 - 12:34:33 AM
Document(s) archivé(s) le : Friday, April 29, 2011 - 3:19:33 AM


Version validated by the jury (STAR)


  • HAL Id : tel-00560535, version 1



David Lanneau. Rôle des protéines de choc thermique HSP90 et HSP70 dans la différenciation macrophagique. Sciences du Vivant [q-bio]. Université de Bourgogne, 2010. Français. <NNT : 2010DIJOS007>. <tel-00560535>




Record views


Document downloads