Dataset Information

Substrates of human TRAP complex

ABSTRACT: While the Sec61-complex in the membrane of the human endoplasmic reticulum facilitates translocation of all precursor polypeptides with amino-terminal signal peptides or transmembrane helices, the Sec61-associated translocon-associated protein (TRAP)-complex supports translocation of only a subset of precursor polypeptides, i.e. in a substrate-specific manner. To characterize TRAP-dependent precursors, we combined siRNA-mediated TRAP depletion in HeLa cells, label-free quantitative proteomics, and differential expression analysis. Sec61 served as a positive control

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Nagarjuna Nagaraj

LAB HEAD: Prof. Dr. Richard Zimmermann

PROVIDER: PXD008178 | Pride | 2018-08-09

REPOSITORIES: Pride

ACCESS DATA

Publications

Proteomics reveals signal peptide features determining the client specificity in human TRAP-dependent ER protein import.

Nguyen Duy D Stutz Regine R Schorr Stefan S Lang Sven S Pfeffer Stefan S Freeze Hudson H HH Förster Friedrich F Helms Volkhard V Dudek Johanna J Zimmermann Richard R

Nature communications 20180914 1

In mammalian cells, one-third of all polypeptides are transported into or across the ER membrane via the Sec61 channel. While the Sec61 complex facilitates translocation of all polypeptides with amino-terminal signal peptides (SP) or transmembrane helices, the Sec61-auxiliary translocon-associated protein (TRAP) complex supports translocation of only a subset of precursors. To characterize determinants of TRAP substrate specificity, we here systematically identify TRAP-dependent precursors by an ...[more]

PMID: 30217974

Similar Datasets

Project description:Glioblastoma multiforme (GBM) is the most prevalent type of adult brain tumor, and one of the deadliest tumors known to mankind. The genetic understanding of GBM is, however, limited, and the molecular mechanisms which facilitate GBM cell survival and growth within the tumor microenvironment are largely unknown. We applied digital karyotyping and single nucleotide polymorphism (SNP) arrays to screen for copy number changes in GBM samples and found that the most frequently amplified region is at chromosome 7p11.2. The high resolution of digital karyotyping and SNP arrays permits the precise delineation of amplicon boundaries and has enabled identification of the minimal region of amplification at 7p11.2, which contains two genes, EGFR and SEC61?. SEC61? encodes a subunit of a heterotrimeric protein channel located in the endoplasmic reticulum (ER). In addition to its high frequency of gene amplification in GBMs, SEC61? is also remarkably overexpressed in 77% of GBMs, but not in lower-grade gliomas. The siRNA-mediated knockdown of SEC61? expression in tumor cells led to growth suppression and apoptosis. Furthermore, we showed that pharmacological ER stress agents induce SEC61? expression in GBM cells. Together, these results indicate that aberrant expression of SEC61? serves significant roles in GBM cell survival, likely via a mechanism that is involved in the cytoprotective ER stress-adaptive response to the tumor microenvironment. hSETD1A was silenced in HCT116 cells using retrovirus harboring shRNA specifically against the hSETD1A gene. 10?g RNA was reverse transcribed to cDNA using the Applied Biosystems High Capacity cDNA Reverse Transcription kit according to the manufacturer’s instructions (Applied Biosystems). cDNA products were treated with 100ng RNaseA and then purified using the Qiagen PCR purification kit according to the manufacturer’s instructions. NimbleGen Human Gene Expression array was purchased from Roche Applied Sciences. cDNA samples were labeled, Cy3 hybridized, and processed at the FSU NimbleGen Microarray Facility at Florida State University.

Dataset Information

Substrates of human TRAP complex

Publications

Proteomics reveals signal peptide features determining the client specificity in human TRAP-dependent ER protein import.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure