A Two-Hybrid Screen of the Yeast Proteome for Hsp90 Interactors Uncovers a Novel Hsp90 Chaperone Requirement in the Activity of a Stress-Activated Mitogen-Activated Protein Kinase, Slt2p (Mpk1p)

Millson, Stefan H.; Truman, Andrew W.; King, Victoria; Prodromou, Chrisostomos; Pearl, Laurence H.; Piper, Peter W.

doi:10.1128/ec.4.5.849-860.2005

Cited by 163 publications

(215 citation statements)

References 73 publications

(78 reference statements)

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“…We now add Pf Hsp90 to the list of heterologous Hsp90's that can apparently fulfill at least the essential functions of Hsp90 in yeast for vegetative growth in standard growth media. Whatever these functions are exactly, they must include many of the ones that have been highlighted by global biochemical and genetic analyses of Hsp90 interactions in this organism [25][26][27][28]. The results of these genetic complementation experiments underscore the high evolutionary conservation of Hsp90 [29][30][31][32].…”

Section: Discussionmentioning

confidence: 99%

The complementation of yeast with human or Plasmodium falciparum Hsp90 confers differential inhibitor sensitivities

Wider

Péli-Gulli

Briand

et al. 2009

Molecular and Biochemical Parasitology

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Developing novel drugs against the unicellular parasite Plasmodium is complicated by the paucity of simple screening systems. Heat-shock proteins are an essential class of proteins for the parasite's cyclical life style between different cellular milieus and temperatures. The molecular chaperone Hsp90 assists a large variety of proteins, but its supporting functions for many proteins that are important for cancer have made it into a well-studied drug target. With a better understanding of the differences between Hsp90 of the malarial parasite and Hsp90 of its human host, new therapeutic options might become available. We have generated a set of isogenic strains of the budding yeast Saccharomyces cerevisiae where the essential yeast Hsp90 proteins have been replaced with either of the two human cytosolic isoforms Hsp90α or Hsp90β, or with Hsp90 from Plasmodium falciparum (Pf). All strains express large amounts of the Flag-tagged Hsp90 proteins and are viable. Even though the strain with Pf Hsp90 grows more poorly, it provides a tool to reconstitute additional aspects of the parasite Hsp90 complex and its interactions with substrates in yeast as a living test tube. Upon exposure of the set of Hsp90 test strains to the two Hsp90 inhibitors radicicol (Rd) and geldanamycin (GA), we found that the strain with Pf Hsp90 is relatively more sensitive to GA than to Rd compared to the strains with human Hsp90's. This indicates that this set of yeast strains could be used to screen for new Pf Hsp90 inhibitors with a wider therapeutic window

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Section: Discussionmentioning

confidence: 99%

The complementation of yeast with human or Plasmodium falciparum Hsp90 confers differential inhibitor sensitivities

Wider

Péli-Gulli

Briand

et al. 2009

Molecular and Biochemical Parasitology

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“…This mutant has been shown to strengthen interactions between HSP90 and substrate peptides in vitro and is predicted to function as a kinetic trap to detect interactions with client proteins and co-chaperones in vivo. Indeed, the analogous mutation in yeast was previously shown to strengthen the interaction between the yeast p23 homolog Sba1p and the yeast HSP90, an interaction known to be positively influenced by ATP (40). Therefore, HSP90 E47A is predicted to serve as a suitable probe in this system to identify HSP90 interactions that are stabilized by excess ATP.…”

Section: Identification Of Novel Hsp90 Interactions and Those Influenmentioning

confidence: 99%

A Proteomic Investigation of Ligand-dependent HSP90 Complexes Reveals CHORDC1 as a Novel ADP-dependent HSP90-interacting Protein

Gano

Simon

2010

Molecular & Cellular Proteomics

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Structural studies of the chaperone HSP90 have revealed that nucleotide and drug ligands induce several distinct conformational states; however, little is known how these conformations affect interactions with co-chaperones and client proteins. Here we use tandem affinity purification and LC-MS/MS to investigate the proteome-wide effects of ATP, ADP, and geldanamycin on the constituents of the human HSP90 interactome. We identified 52 known and novel components of HSP90 complexes that are regulated by these ligands, including several co-chaperones. Interestingly, our results also show that geldanamycin treatment causes HSP90 complexes to become significantly enriched for core transcription machinery, suggesting that HSP90 inhibition may have broad based effects on transcription and RNA processing. We further characterized a novel ADP-dependent HSP90 interaction with the cysteine-and histidine-rich domain (CHORD)-containing protein CHORDC1. We show that this interaction is stimulated by high ADP:ATP ratios in cell lysates and in vitro with purified recombinant proteins. Furthermore, we demonstrate that this interaction is dependent upon the ability of HSP90 to bind nucleotides and requires the presence of a linker region between the CHORD domains in CHORDC1. Together these findings suggest that the HSP90 interactome is dynamic with respect to nucleotide and drug ligands and that pharmacological inhibition of HSP90 may stimulate the formation of specific complexes. Molecular & Cellular Proteomics 9:255-270, 2010.

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“…However, several recent studies have focused on identification of Hsp90 partners using large-scale proteomic approaches, such as immunoprecipitation, immobilization of the Cdomain of Hsp90α, genome-wide two-hybrid screens, and proteome analysis of tumor cells subjected to treatment with Hsp90 inhibitor (Falsone et al 2005;Millson et al 2005;Zhao et al 2005;McClellan et al 2007;Schumacher et al 2007;Te et al 2007). These studies have led to the identification of a number of novel Hsp90-interacting partners, including cochaperones and client proteins, and suggested several previously unknown functions of Hsp90, for example, cellular transport, cytokinesis, and epigenetic gene regulation.…”

Section: Introductionmentioning

confidence: 99%

Novel Hsp90 partners discovered using complementary proteomic approaches

Tsaytler

Krijgsveld

Goerdayal

et al. 2009

Cell Stress and Chaperones

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Hsp90 is an essential eukaryotic molecular chaperone that stabilizes a large set of client proteins, many of which are involved in various cellular signaling pathways. The current list of Hsp90 interactors comprises about 200 proteins and this number is growing steadily. In this paper, we report on the application of three complementary proteomic approaches directed towards identification of novel proteins that interact with Hsp90. These methods are coimmunoprecipitation, pull down with biotinylated geldanamycin, and immobilization of Hsp90β on sepharose. In all, this study led to the identification of 42 proteins, including 18 proteins that had not been previously characterized as Hsp90 interactors. These novel Hsp90 partners not only represent abundant protein species, but several proteins were identified at low levels, among which signaling kinase Cdk3 and putative transcription factor tripartite motif-containing protein 29. Identification of tetratricopeptide-repeat-containing mitochondrial import receptor protein Tom34 suggests the involvement of Hsp90 in the early steps of translocation of mitochondrial preproteins. Taken together, our data expand the knowledge of the Hsp90 interactome and provide a further step in our understanding of the Hsp90 chaperone system.

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A Two-Hybrid Screen of the Yeast Proteome for Hsp90 Interactors Uncovers a Novel Hsp90 Chaperone Requirement in the Activity of a Stress-Activated Mitogen-Activated Protein Kinase, Slt2p (Mpk1p)

Cited by 163 publications

References 73 publications

The complementation of yeast with human or Plasmodium falciparum Hsp90 confers differential inhibitor sensitivities

The complementation of yeast with human or Plasmodium falciparum Hsp90 confers differential inhibitor sensitivities

A Proteomic Investigation of Ligand-dependent HSP90 Complexes Reveals CHORDC1 as a Novel ADP-dependent HSP90-interacting Protein

Novel Hsp90 partners discovered using complementary proteomic approaches

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