Dataset Information

CHN-1 and CHN-1 L141R interacting proteins

ABSTRACT: Proteostasis is achieved by quality control pathways that support the generation of correctly folded proteins, prevent protein misfolding and remove toxic proteins. The quality control E3 ligase CHIP ubiquitylates damaged proteins consigned by chaperone partners for disposal through the endo-lysosomal pathway, proteasomal degradation, or autophagy. Additionally, CHIP has been reported to modulate essential signaling pathways by precisely delivering a myriad of native proteins to destined fates. We aimed at understanding the substrate specificity and processivity through a “structure to function” approach, by examining the modeled 3D structure of the C. elegans ortholog of CHIP, CHN-1, based on the reported structure of murine CHIP. Using different model organisms and various genetic and biochemical analyses, we demonstrate that monomeric CHN-1/CHIP has preserved ubiquitylation activity and promotes longevity via the IIS pathway. The lack of monomer results in premature aging and neurodegenerative disorder. Our data reveal that autoubiquitylation plays an important role in the alteration between monomer and dimer. Together, the conserved dimer-monomer transition provides a molecular switch regulating CHIP activity in response to proteotoxic stress and aging.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Caenorhabditis Elegans

SUBMITTER: Thorsten Hoppe

LAB HEAD: Thorsten Hoppe

PROVIDER: PXD025375 | Pride | 2022-08-25

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20190416_Q4_JK_ColID_768_ProjID_1724_Hoppe_sample_01.raw	Raw
	20190416_Q4_JK_ColID_768_ProjID_1724_Hoppe_sample_02.raw	Raw
	20190416_Q4_JK_ColID_768_ProjID_1724_Hoppe_sample_03.raw	Raw
	20190416_Q4_JK_ColID_768_ProjID_1724_Hoppe_sample_04.raw	Raw
	20190416_Q4_JK_ColID_768_ProjID_1724_Hoppe_sample_05.raw	Raw

Items per page:

1 - 5 of 14

Similar Datasets

Project description:This project includes two proximity labeling experiments, one to establish the vicinity of UNC-45 under non-stress conditions and one to examine changes in the vicinity of UNC-45 under optogenetically induced mechanical muscle stress. The first non-stress experiment consists of 15 samples after biotin-streptavidin pull-down in 5 replicates of 3 conditions (3 transgenic C. elegans strains). Transgenic C. elegans strains expressing the mutant BirA biotin ligases miniTurbo and TurboID in body wall muscle cells were used to compare a proximity-labeled strain expressing an UNC-45-miniTurbo-2xHA fusion protein in the muscle cells (PP3135 unc-119(ed4)III; hhIs241[unc-54p::unc-45::miniTurbo::2xHA; unc-119(+)]) with a strain expressing the biotin ligase TurboID-2xHA without fusion protein in muscle cells (PP3138 unc-119(ed4)III; hhIs242[unc-54p::TurboID::2xHA; unc-119(+)]) to find transient interactors of the myosin chaperone UNC-45 in muscle. As a control, we included a strain expressing a transgenic UNC-45-FLAG protein in the body wall muscle (PP1017 unc-119(ed4)III; hhIs84[unc-119(+); unc-54p::unc-45::FLAG]). Biotinylated proteins in 2.5 mg lysates of these strains were pulled down with streptavidin sepharose beads and identified by mass spectrometry. The second mechanical stress experiment consists of 10 samples after biotin-streptavidin pull-down in 5 replicates of 2 conditions. A transgenic C. elegans strain expressing the biotin ligase miniTurbo fused to UNC-45 in body wall muscle cells was crossed with a transgenic strain expressing the optogenetic channelrhodopsin mutant ChR2(C128S;H134R)-FLAG in body wall muscle cells (PP3358 hhIs241[unc-54p::unc-45::miniTurbo::2xHA; unc-119(+)]; hhIs251[myo-3p::ChR2(C128S,H134R)-FLAG::unc-54 3'UTR, Cbrunc-119(+)]). The latter transgene is used to contract the worms’ muscles by blue light illumination to induce mechanical muscle stress. The optogenetic channel is activated by adding the cofactor all-trans retinal (ATR) to its food source E. coli OP50. In this experiment, we compared proximity-labeling in the UNC-45-miniTurbo-2xHA expressing strain under conditions with ATR (L+, treatment, contraction) with proximity-labeling in the UNC-45-miniTurbo-2xHA expressing strain under conditions without ATR (L-, control, no contraction) to find transient interactors of the candidate UNC-45 in muscle under mechanical stress. Biotinylated proteins in 2.5 mg of lysates of these strains were pulled down using streptavidin sepharose beads and identified by mass spectrometry.

Dataset Information

CHN-1 and CHN-1 L141R interacting proteins

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets