Dataset Information

Juvenile hormone membrane signaling phosphorylates USP to potentiate 20-hydroxyecdysone action and Yorkie activity in Drosophila

ABSTRACT: Juvenile hormone (JH) antagonizes 20-hydroxyecdysone (20E) signaling mainly through the intracellular receptors Met and Gce in Drosophila. To investigate JH membrane signaling pathway without the interference from JH intracellular signaling, we characterized phosphoproteome profiles of the Met gce double mutant in the absence or presence of JH in both chronic and acute phases. Importantly, JH was identified to phosphorylate USP at Ser35, the PKC phosphorylation site required for the maximal action of 20E through its nuclear receptor complex EcR-USP. Functioning through a potential receptor tyrosine kinase and phospholipase C pathway, JH membrane signaling activated PKC to phosphorylate USP at Ser35. The USPS35A mutant, in which Ser35 was replaced with Ala35 by genome editing, showed attenuated 20E signaling and Yorkie activity, resulting in delayed developmental timing and reduced body size, respectively. Moreover, genetic interaction experiments confirmed that JH lied upstream of PKC-mediated USP phosphorylation at Ser35. This study propose a novel model of JH membrane signaling that exhibits crosstalk with different pathways simultaneously to regulate distinct developmental processes.

ORGANISM(S): Drosophila Melanogaster

SUBMITTER: Sheng Li

PROVIDER: PXD021644 | iProX | Wed Sep 23 00:00:00 BST 2020

REPOSITORIES: iProX

ACCESS DATA

Publications

Juvenile hormone membrane signaling phosphorylates USP and thus potentiates 20-hydroxyecdysone action in Drosophila.

Gao Yue Y Liu Suning S Jia Qiangqiang Q Wu Lixian L Yuan Dongwei D Li Emma Y EY Feng Qili Q Wang Guirong G Palli Subba R SR Wang Jian J Li Sheng S

Science bulletin 20210624 2

Juvenile hormone (JH) and 20-hydroxyecdysone (20E) coordinately regulate development and metamorphosis in insects. Two JH intracellular receptors, methoprene-tolerant (Met) and germ-cell expressed (Gce), have been identified in the fruit fly Drosophila melanogaster. To investigate JH membrane signaling pathway without the interference from JH intracellular signaling, we characterized phosphoproteome profiles of the Met gce double mutant in the absence or presence of JH in both chronic and acute ...[more]

PMID: 36546012

Similar Datasets

Project description:MET, the receptor for hepatocyte growth factor (HGF), is involved in wide variety of biological events and its aberrant activation is implicated in the pathogenesis of cancer. Especially, MET signaling is associated with resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), key drugs in therapy of non-small cell lung cancer. MET has 11 potential N-glycosylation sites, however, the site-specific roles of N-glycans have not been elucidated. Here, we report that N-glycans regulate proteolytic processing of MET and HGF induced MET signaling, site specifically. N-glycosylation inhibitors suppressed processing and trafficking of endogenous MET in H1975 and EBC-1 lung cancer cells and exogenous MET in CHO cells. We purified the recombinant extracellular domain of human MET and determined the site-specific N-glycan structures and occupancy using nanoflow liquid chromatography-electrospray ionization mass spectrometry. The results indicated that most sites were fully glycosylated and the dominant population were complex-type which were rich in sialic acids and core fucoses. To examine the effects of N-glycan deletion of MET, we prepared endogenous-MET-knockout Flp-In CHO cells and transfected with series of N-glycan deletion mutants of MET. It was found that several N-glycans are implicated in processing of MET. It was also suggested that the N-glycans of -subunit of MET positively regulate HGF signaling, and the N-glycans of β-subunit negatively regulate HGF signaling. In all-N-glycan deletion mutant, processing and signaling were significantly suppressed. It was observed that the cell proliferation rate was suppressed in all-N-glycan deletion mutant, although the cell surface expression levels of MET were not completely diminished. We examined the HGF biding affinity for the recombinant extracellular domain of MET and found that peptide N-glycosidase F treatment did not affect the ligand binding affinity. It was suggested that N-glycans of MET affect the status and the function of the receptor site-specifically.

Dataset Information

Juvenile hormone membrane signaling phosphorylates USP to potentiate 20-hydroxyecdysone action and Yorkie activity in Drosophila

Publications

Juvenile hormone membrane signaling phosphorylates USP and thus potentiates 20-hydroxyecdysone action in Drosophila.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure