PXD032168

PXD032168 is an original dataset announced via ProteomeXchange.

Title	The pseudokinase NRBP1 activates Rac/Cdc42 via P-Rex1 to drive oncogenic signalling in triple negative breast cancer
Description	In an effort to identify oncogenic protein kinase drivers in triple negative breast cancer (TNBC), we undertook mass spectrometry (MS)-based proteomic profiling across a large panel of TNBC cell lines. This revealed marked variation in expression of the multidomain pseudokinase Nuclear Receptor Binding Protein 1 (NRBP1). Interrogation of publicly-available data determined that NRBP1 gene expression is higher in TNBC than in luminal breast cancers and positively associates with poor prognosis in TNBC patients. Gain and loss of function experiments characterized NRBP1 as a positive regulator of TNBC cell migration and invasion. In addition, NRBP1 knockdown reduced colony formation in vitro and moreover, markedly impaired growth of TNBC orthotopic xenografts as well as lung colonization in an experimental metastasis model. To interrogate the signalling mechanism of NRBP1, we applied BioID/MS profiling, identifying P-Rex1, a guanine nucleotide exchange factor for Rac1 and Cdc42, as a NRBP1 binding partner. Importantly, NRBP1 overexpression enhanced levels of GTP-bound Rac1 and Cdc42 in a P-Rex1-dependent manner, while NRBP1 knockdown reduced their activation. In addition, NRBP1 associated with P-Rex1, Rac1 and Cdc42, suggesting a scaffolding function for this pseudokinase. NRBP1-mediated promotion of cell migration and invasion was P-Rex1-dependent, while constitutively-active Cdc42 wholly/or partially rescued the effect of NRBP1 knockdown on cell migration/invasion and colony formation, respectively. Generation of reactive oxygen species via a NRBP1/P-Rex1 pathway was implicated in these oncogenic roles of NRBP1. Overall, these findings define a new function for NRBP1 and a novel oncogenic signalling pathway in TNBC that may be amenable to therapeutic intervention.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:55:22.110.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ralf Schittenhelm
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2022-03-09 02:35:56	ID requested
1	2023-05-04 20:36:13	announced
⏵ 2	2023-11-14 08:55:22	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: G proteins, proteomics,signal transduction, scaffold proteins

Roger Daly
contact affiliation	Cancer Program, Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Melbourne, VIC, 3800, Australia
contact email	roger.daly@monash.edu
lab head
Ralf Schittenhelm
contact affiliation	Monash University
contact email	ralf.schittenhelm@monash.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/05/PXD032168

PRIDE project URI

• PRIDE
  1. PXD032168
     1. Label: PRIDE project
     2. Name: The pseudokinase NRBP1 activates Rac/Cdc42 via P-Rex1 to drive oncogenic signalling in triple negative breast cancer