Structural Basis for Phosphorylation-Dependent Recruitment of Tel2 to Hsp90 by Pih1

Pal, Mohinder; Morgan, Rhodri M. L.; Phelps, Sarah; Roe, S. Mark; Parry-Morris, Sarah; Downs, Jessica A.; Polier, Sigrun; Pearl, Laurence H.; Prodromou, Chrisostomos

doi:10.1016/j.str.2014.04.001

Cited by 91 publications

(208 citation statements)

References 38 publications

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“…In SMG-1 the a-solenoid extends away from the kinase domain, and the restriction to the active site might be carried out by the 1200-residue long kinase domain insertion instead (Figures 3d, 4a). The conformational maturation of PIKKs is tightly controlled by a supercomplex composed of Hsp90 and a cochaperone system composed of TTT (TEL2-TTI1-TTI2) and R2TP (RUVBL1-RUVBL2-RPAP3-PIH1D1) [50,51], with the R2TP complex acting as a bridge between Hsp90 and the TTT complex [52]. This chaperone assembly most likely binds to the helical repeats of newly synthesized PIKKs.…”

Section: The Pikk A-solenoidmentioning

confidence: 99%

PIKKs — the solenoid nest where partners and kinases meet

Baretić

Williams

2014

Current Opinion in Structural Biology

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Section: The Pikk A-solenoidmentioning

confidence: 99%

PIKKs — the solenoid nest where partners and kinases meet

Baretić

Williams

2014

Current Opinion in Structural Biology

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“…Similar examples illustrate an adaptor/recruiter function for Pih1 in recruiting the Hsp90 chaperone to other complexes. These include the recruitment of Hsp90 to the yeast box C/D snoRNP assembly via an interaction between Pih1 and Nop58 (4) and the activation of PIKK signaling by recruitment of Hsp90 to PIKK via an interaction between Pih1 and the PIKK subunit Tel2 (9).…”

mentioning

confidence: 99%

Proteomic and Genomic Analyses of the Rvb1 and Rvb2 Interaction Network upon Deletion of R2TP Complex Components

Lakshminarasimhan

Boanca

Banks

et al. 2016

Molecular & Cellular Proteomics

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The highly conserved yeast R2TP complex, consisting of Rvb1, Rvb2, Pih1, and Tah1, participates in diverse cellular processes ranging from assembly of protein complexes to apoptosis. Rvb1 and Rvb2 are closely related proteins belonging to the AAA؉ superfamily and are essential for cell survival. Although Rvbs have been shown to be associated with various protein complexes including the Ino80 and Swr1chromatin remodeling complexes, we performed a systematic quantitative proteomic analysis of their associated proteins and identified two additional complexes that associate with Rvb1 and Rvb2: the chaperonin-containing T-complex and the 19S regulatory particle of the proteasome complex. We also analyzed Rvb1 and Rvb2 purified from yeast strains devoid of PIH1 and TAH1. These analyses revealed that both Rvb1 and Rvb2 still associated with Hsp90 and were highly enriched with RNA polymerase II complex components. Our analyses also revealed that both Rvb1 and Rvb2 were recruited to the Ino80 and Swr1 chromatin remodeling complexes even in the absence of Pih1 and Tah1 proteins. Using further biochemical analysis, we showed that Rvb1 and Rvb2 directly interacted with Hsp90 as well as with the RNA polymerase II complex. RNA-Seq analysis of the deletion strains compared with the wild-type strains revealed an up-regulation of ribosome biogenesis and ribonucleoprotein complex biogenesis genes, down-regulation of response to abiotic stimulus genes, and downregulation of response to temperature stimulus genes. A Gene Ontology analysis of the 80 proteins whose protein associations were altered in the PIH1 or TAH1 deletion strains found ribonucleoprotein complex proteins to be the most enriched category. This suggests an important function of the R2TP complex in ribonucleoprotein complex biogenesis at both the proteomic and genomic levels. The Saccharomyces cerevisiae R2TP complex consists of Rvb1, Rvb2, Pih1 (protein interacting with Hsp90) and Tah1 (TPR (tetratricopeptide repeat)-containing protein associated with Hsp90) proteins. Although, the R2TP complex was initially discovered in yeast as an Hsp90-associated complex (1), R2TP subunits are conserved in higher eukaryotes and the R2TP complex was recently identified and purified from human cells (2). The R2TP complex has been implicated in various cellular processes such as assembly of small nucleolar ribonuclearprotein (snoRNP) 1 (small nucleolar ribonucleoprotein) complex (3, 4), RNAPII (RNA polymerase II) complex (5), apoptosis (6), and PIKK (phosphatidylinositol 3-kinase-related protein kinases) signaling (7).

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“…4B) were able to bind to Rpn8. These data suggest that the mechanism of Pih1/Rpn8 binding is different from the one between Pih1 and Tah1, because Tah1 binds to the CS domain of Pih1 (20), and the fragment Pih1(282-344) alone is not able to bind Tah1 (8).…”

Section: Pih1 C-terminal Fragment Triggers Proteasome-dependentmentioning

confidence: 87%

“…The atomic structures of the Pih1 domain have been solved for both yeast and human homologues (19,20). The Pih1 domain adopts an unusual ␤␤␣␤␤␤␣␣ topology.…”

Section: The R2tpmentioning

confidence: 99%

“…The yeast Pih1 C-terminal fragment contains a CS domain, which also appears in other proteins, such as HSP90 co-chaperones p23 and Sgt1 (21,22), and interacts with HSP90 directly (23). The Tah1 C-terminal fragment forms a constitutive interaction with the Pih1 CS domain through main chain interactions, in one way by forming ␤-strand to extend the ␤-sheet of the Pih1 CS domain and in the other way by bridging the edges of the two ␤-sheets of the CS domain (20). The tight interaction between Tah1 and Pih1 has also been observed when they are co-expressed in Escherichia coli (24).…”

Section: The R2tpmentioning

confidence: 99%

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The Proteasome Subunit Rpn8 Interacts with the Small Nucleolar RNA Protein (snoRNP) Assembly Protein Pih1 and Mediates Its Ubiquitin-independent Degradation in Saccharomyces cerevisiae

Paci

Liu

Zhang

et al. 2016

Journal of Biological Chemistry

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Pih1 is a scaffold protein of the Rvb1-Rvb2-Tah1-Pih1 (R2TP) protein complex, which is conserved in fungi and animals. The chaperone-like activity of the R2TP complex has been implicated in the assembly of multiple protein complexes, such as the small nucleolar RNA protein complex. However, the mechanism of the R2TP complex activity in vivo and the assembly of the complex itself are still largely unknown. Pih1 is an unstable protein and tends to aggregate when expressed alone. The C-terminal fragment of Pih1 contains multiple destabilization factors and acts as a degron when fused to other proteins. In this study, we investigated Pih1 interactors and identified a specific interaction between Pih1 and the proteasome subunit Rpn8 in yeast Saccharomyces cerevisiae when HSP90 co-chaperone Tah1 is depleted. By analyzing truncation mutants, we identified that the C-terminal 30 amino acids of Rpn8 are sufficient for the binding to Pih1 C terminus. With in vitro and in vivo degradation assays, we showed that the Pih1 C-terminal fragment Pih1(282-344) is able to induce a ubiquitin-independent degradation of GFP. Additionally, we demonstrated that truncation of the Rpn8 C-terminal disordered region does not affect proteasome assembly but specifically inhibits the degradation of the GFP-Pih1(282-344) fusion protein in vivo and Pih1 in vitro. We propose that Pih1 is a ubiquitin-independent proteasome substrate, and the direct interaction with Rpn8 C terminus mediates its proteasomal degradation. The R2TP2 complex, which is composed of Rvb1, Rvb2, Tah1, and Pih1, was first discovered in baking yeast (1) and is required for box C/D small nucleolar RNA protein (snoRNP) complex assembly and thus for the ribosomal RNA biogenesis (2). R2TP is a conserved protein complex and has also been identified in mammalian cells (3-5), fruit fly (6), plasmodium (7), and many other fungi species (8), whereas it is absent in higher plants (9). There are more than 70 different snoRNP complexes that are directly involved in ribosomal RNA processing, a critical step for ribosome assembly that requires a total of more than 200 assembly factors (10). The assembly of R2TP complex in vivo is also a highly dynamic process and is regulated by the molecular chaperone HSP90 and nutrition status (11). However, the mechanism by which R2TP controls box C/D snoRNP complex assembly is still largely unknown.Pih1, the scaffold protein within the R2TP complex, mediates the interaction between Tah1 and Rvb1-Rvb2 and subsequently controls the biogenesis of box C/D snoRNP (2, 3). Tah1 recruits HSP90 and protects Pih1 from degradation in vivo (12, 13). Pih1 interacts directly with Nop58, one of the core protein subunits in box C/D snoRNP (11) and affects its stability (14). Pih1 also interacts with Rsa1, another snoRNP assembly factor through which the R2TP complex may interact with the snoRNP core protein Snu13 (15). Similar to the role of Tah1 in protecting Pih1, a small protein, Hit1, was also identified to interact with and protect Rsa1, thus revealing a complex...

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Structural Basis for Phosphorylation-Dependent Recruitment of Tel2 to Hsp90 by Pih1

Cited by 91 publications

References 38 publications

PIKKs — the solenoid nest where partners and kinases meet

PIKKs — the solenoid nest where partners and kinases meet

Proteomic and Genomic Analyses of the Rvb1 and Rvb2 Interaction Network upon Deletion of R2TP Complex Components

The Proteasome Subunit Rpn8 Interacts with the Small Nucleolar RNA Protein (snoRNP) Assembly Protein Pih1 and Mediates Its Ubiquitin-independent Degradation in Saccharomyces cerevisiae

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