Tsc1 co‐chaperone inhibits Hsp90 chaperone function

Slides:

Advertisements

Similar presentations

Supplementary Figures

Advertisements

Exogenous B56γ3 WT F395C WT F395C IgG Vinc Endogenous B56γ2,3 HA-IPInput Figure S1 HA-B56γ3 is unable to interact with endogenous B56γ. Lysates of U2OS.

Loss of Hdac3 impairs Ar signaling in mouse prostate tissues and has no effect on apoptosis Loss of Hdac3 impairs Ar signaling in mouse prostate tissues.

Resveratrol inhibits the transactivation potential of the RelA/p65 protein. Resveratrol inhibits the transactivation potential of the RelA/p65 protein.

Cdc42.GTP directly activates PAK4.

Volume 11, Issue 2, Pages (April 2015)

JNK is activated by Wnt5a.

SERCA2 is a functional adaptor for the alternative toll‐like receptor 9 (TLR9) signalling in cardiomyocytes Co‐immunoprecipitated TLR9 with SERCA2 antibody.

The N‐terminus and SH2 domain of SOCS‐1 contribute to inhibition of JAK1 and JAK2 kinase activity. The N‐terminus and SH2 domain of SOCS‐1 contribute to.

(A) Left panel: rate of H2O2 release per minute was analysed by calculating the linear slope of data obtained in Supplementary Figure S1A. (A) Left panel:

AO domain of LSD1 is responsible for its interaction with Snail1.

Volume 36, Issue 5, Pages (December 2009)

SkMLCK physically interacts with and phosphorylates MEF2C in vivo and in vitro. skMLCK physically interacts with and phosphorylates MEF2C in vivo and in.

HDAC1 is an integral component of the Microprocessor complex and modifies its affinity to primary miRNA transcripts. HDAC1 is an integral component of.

Volume 5, Issue 4, Pages (April 2004)

PGAM5 associates with and activates ASK1.

Volume 18, Issue 1, Pages (January 2008)

Ubiquitination of Drp1 by MARCH‐V.

NoRC regulates telomeric heterochromatin.

Volume 21, Issue 7, Pages (November 2017)

Expression levels of the transgenic DIA1(R1204X) mutant in the inner ear of TG mice Expression levels of the transgenic DIA1(R1204X) mutant in the inner.

Mutations affecting HOIP–UBA and HOIL‐1L–UBL interaction that compromise NF‐κB activation. Mutations affecting HOIP–UBA and HOIL‐1L–UBL interaction that.

Rpn11p is recruited to the tombusvirus replicase in yeast.

Attenuation of AML1-ETO cellular dysregulation correlates with increased leukemogenic potential by Russell C. DeKelver, Ming Yan, Eun-Young Ahn, Wei-Jong.

HS induces sustained H3K4me3 and H3K4me2 methylation at HSP22

MCPH1 interacts with E2F1. MCPH1 interacts with E2F1. (A,B) HEK293 cells were left untreated (No tx) or treated with 300 ng/ml NCS for 4 h or 2 μM adriamycin.

Reactive oxygen species inhibit miR‐199a and miR‐125b expression levels in vitro and in vivo. Reactive oxygen species inhibit miR‐199a and miR‐125b expression.

NRF2 Is a Major Target of ARF in p53-Independent Tumor Suppression

Palbociclib activates the proteasome indirectly and reduces the association of ECM29 with the proteasome Palbociclib activates the proteasome indirectly.

DDX3 is required for TBK1/IKKε‐dependent promoters and is recruited to IKKε. DDX3 is required for TBK1/IKKε‐dependent promoters and is recruited to IKKε.

KLK7 is involved in the Aβ degradation activity

Expression of PD‐1 on PBMC and malignant exudate T cells

The de‐ubiquitinase OTUB1 interacts with the RAS GTPases

Effect of HUWE1 on MYC and MIZ1 complexes Immunoblot documenting levels of MYC and MIZ1 upon HUWE1 overexpression. Effect of HUWE1 on MYC and MIZ1 complexes.

WDR11 forms a tertiary complex with EMX1 and GLI3

Volume 6, Issue 6, Pages (March 2014)

Volume 12, Issue 5, Pages (November 2003)

HDAC5, a Key Component in Temporal Regulation of p53-Mediated Transactivation in Response to Genotoxic Stress Nirmalya Sen, Rajni Kumari, Manika Indrajit.

Volume 27, Issue 6, Pages (September 2007)

MED12 association with ncRNA-a is disrupted by loss of CARM1 and TDRD3

RIG‐I (N) and LRRC25 interact with ISG15 in an unconjugated manner, related to Figs 6 and 7 RIG‐I (N) and LRRC25 interact with ISG15 in an unconjugated.

USP26 binds to and deubiquitinates SMAD7

Javier Sánchez-Pozo, Alexander J. Baker-Williams, Mark R

NDP52 promotes CHIKV infection in a species‐specific manner.

Volume 41, Issue 6, Pages (March 2011)

LRRC25 inhibits RLR‐mediated type I IFN signaling pathway

Volume 22, Issue 2, Pages (January 2018)

Protein tyrosine kinase 7 is involved in Wnt/β‐catenin canonical signalling. Protein tyrosine kinase 7 is involved in Wnt/β‐catenin canonical signalling.

The CHCH domain is necessary for mitochondrial import of CHCHD10

HDAC11 regulates CCL2 expression by recruiting PU.1.

Mass spectrometric validation of DLG5 as a CDKL5 substrate

Volume 70, Issue 3, Pages e6 (May 2018)

ATAD1 physically interacts with the TA protein, GOS28, and is required to limit the level of mislocalized GOS28 on mitochondria in mammals Human dermal.

Yeast Msp1 and human ATAD1 are required to limit the level of mitochondrially mislocalized TA proteins Pex15 and PEX26, respectively Representative images.

Volume 49, Issue 6, Pages (March 2013)

Hua Gao, Yue Sun, Yalan Wu, Bing Luan, Yaya Wang, Bin Qu, Gang Pei

NLK positively regulates YAP activity in physiological settings

Abraxas binds to BRCT–PALB2(L21P) and is involved in the recruitment of this fusion protein or of endogenous PALB2 to sites of DNA damage. Abraxas binds.

Volume 21, Issue 9, Pages (November 2017)

Mutant MFN2 proteins have normal half-lives and are properly localized to mitochondria. Mutant MFN2 proteins have normal half-lives and are properly localized.

Binding of p38α and p38β2 to MAP kinase kinases

Chk1 and 14‐3‐3 proteins inhibit atypical E2Fs to prevent a permanent cell cycle arrest Experimental procedures of the SILAC assay to identify binding.

Fig. 1 SAMTOR interacts with GATOR1 and KICSTOR.

Tumor suppressor Tsc1 is a new Hsp90 co‐chaperone that facilitates folding of kinase and non‐kinase clients Tsc1 was immunoprecipitated from HEK293 cell.

Gain-of-function conferred by RNF31 SNPs

Volume 28, Issue 7, Pages e6 (August 2019)

Fig. 3 DUB proteins act at different levels of type I IFN pathways.

OPCML directly interacts with the extracellular domain of EPHA2, FGFR1, and HER2, a prerequisite for downregulation. OPCML directly interacts with the.

FIH hydroxylates G9a and GLP at asparagine residues.

Volume 25, Issue 1, Pages (January 2007)

Presentation transcript:

Tsc1 co‐chaperone inhibits Hsp90 chaperone function Tsc1 co‐chaperone inhibits Hsp90 chaperone function Tsc1 was immunoprecipitated from HEK293 cell lysates using anti‐Tsc1 antibody and immunoblotted with indicated antibodies to confirm chaperone, co‐chaperone, and client protein interaction. IgG was used as control.FLAG‐tagged Hsp90α N‐, M‐, and C‐domains were transiently expressed and immunoprecipitated from HEK293 cells. Co‐immunoprecipitation (Co‐IP) of endogenous Tsc1 and Tsc2 was examined by immunoblotting. Empty vector (EV) was used as a control.Hsp90α‐FLAG, Hsp90β‐FLAG, and their deleted TPR domain‐binding site constructs were transiently expressed and immunoprecipitated from HEK293 cells. Interaction of Tsc1 was assessed by immunoblotting. HOP was used as a positive control. Empty vector (EV) was used as a control.FLAG‐tagged Tsc1 domains were transiently expressed and isolated from HEK293 cells. Co‐IP of endogenous Hsp70 and Hsp90 was assessed by immunoblotting. Empty vector (EV) was used as a control.Tsc1‐D‐HA was co‐expressed with FLAG‐tagged Hsp90α N‐, M‐, or C‐ domains in HEK293 cells. Different domains of Hsp90α‐FLAG were immunoprecipitated, and Tsc1‐D‐HA interaction was assessed by immunoblotting. Empty vector (EV) was used as a control.Bacterially expressed and purified Tsc1‐D‐His6 and Hsp90α binding affinity in the presence of AMPPNP, ADP, and GB were measured by fluorescence anisotropy. The error bars represent mean ± SD of three independent experiments.ATPase activity of Hsp90α in vitro. Inhibitory effects of purified Tsc1‐D‐His6 on the ATPase activity of Hsp90α are shown. All the data represent mean ± SD of three biological replicates. A Student's t‐test was performed to assess statistical significance (**P < 0.005, ****P < 0.0001).Hsp90 was isolated and ATPase activity was measured from five Tsc1GFAPCKO mice with conditional inactivation of the TSC1 gene predominantly in glia and five Tsc1flox/+‐GFAP‐Cre and Tsc1flox/flox littermate controls (Tsc1‐WT). A Student's t‐test was performed to assess statistical significance (**P < 0.005).Tsc1, Hsp90, and client proteins from samples in (H) were examined by immunoblotting. SE (short exposure) and LE (long exposure) of the radiographic film. Source data are available online for this figure. Mark R Woodford et al. EMBO J. 2017;embj.201796700 © as stated in the article, figure or figure legend