PXD049959

PXD049959 is an original dataset announced via ProteomeXchange.

Title	The penetrant chordoid glioma PRKCA mutation is a gain-of-function kinase inactivation eliciting early onset chondrosarcoma in mice
Description	A highly penetrant heterozygous mutation in PRKCA was found recently in chordoid glioma. To establish whether this mutant of PKC was a driver of tumour formation or maintenance in vivo, knock-in mice were generated. The constitutive, heterozygous expression of the Prkca D463H mutant in mice provoked the development of bilateral hindlimb chondrosarcomas and early neonatal mortality. The lack of activity of the D463H mutant was not the trigger of the phenotype as heterozygous knock-in of the related inactivating mutation D463N produced offspring indistinguishable from parental strains. Moreover, this surprising distinction between D463H and D463N mutations did not reflect a selective lack of dominance for the D463N mutation over the wild type (WT) allele since homozygous expression of this mutation was phenotypically WT. In cell culture, the D463H mutant expression, localisation and downregulation behaviours closely mimicked that of the WT, with the D463N mutant displaying some subtle differences. Biochemically, both the D463H and D463N mutants were shown to be inactive, although based on thermal shift assays the D463N mutant had reduced ATP affinity compared to WT or D463H proteins. Using induced transcriptional changes as a functional readout, altered mRNA expression indicates that the D463H acts as a dominant gain-of-function PKC mutant, despite a complete lack of intrinsic kinase activity; transcriptional changes induced by the D463N mutant were distinct from both the WT and D463H mutant. These findings suggest a model in which there is a non-catalytic effector function of PKC which it is presumed operates through protein-protein interactions, as evidenced here by TurboID proximity labelling. The penetrant selection of the specific inactivating D463H mutation in chordoid glioma and the distinction with the D463N mutation indicates that this is conformationally sensitive. The properties characterised suggest this is driven by ATP binding and by inference, in this tissue context, the catalytic activity of this kinase acts as an autoregulatory negative feedback control.
HostingRepository	PRIDE
AnnounceDate	2025-05-29
AnnouncementXML	Submission_2025-05-29_08:05:17.671.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	VERONIQUE CALLEJA
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-02-20 06:17:12	ID requested
⏵ 1	2025-05-29 08:05:18	announced

10.1101/2025.05.28.656646;

submitter keyword: human,PKC alpha, glioma cell line

Peter Parker
contact affiliation	Emeritus Group Leader Protein Phosphorylation Laboratory The Francis Crick Institute 1 Midland Road London NW1 1AT
contact email	peter.parker@crick.ac.uk
lab head
VERONIQUE CALLEJA
contact affiliation	The Francis Crick Institute
contact email	veronique.calleja@crick.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD049959
     1. Label: PRIDE project
     2. Name: The penetrant chordoid glioma PRKCA mutation is a gain-of-function kinase inactivation eliciting early onset chondrosarcoma in mice