PXD035664-1

PXD035664 is an original dataset announced via ProteomeXchange.

Title	Mutational screens highlight glycosylation as a modulator of colony-stimulating factor 3 receptor (CSF3R) activity
Description	The colony-stimulating factor 3 receptor (CSF3R) controls the growth of neutrophils, the most abundant type of white blood cell. In healthy neutrophils, signaling is dependent on CSF3R binding to its ligand, CSF3. A single amino acid mutation in CSF3R, T618I, instead allows for constitutive, ligand-independent cell growth and leads to a rare type of cancer called chronic neutrophilic leukemia (CNL). However, the disease mechanism is not well understood. Here, we investigated why this threonine to isoleucine substitution is the predominant mutation in CNL and how it leads to uncontrolled neutrophil growth. Using protein domain mapping, we demonstrated that the single CSF3R domain containing residue 618 is sufficient for ligand-independent activity. We then applied an unbiased mutational screening strategy focused on this domain and found that activating mutations are enriched at sites normally occupied by asparagine, threonine, and serine residues – the three amino acids which are commonly glycosylated. We confirmed glycosylation at multiple CSF3R residues by mass spectrometry, including the presence of GalNAc and Gal-GalNAc glycans at wild-type threonine 618. Using the same approach applied to other cell surface receptors, we identified an activating mutation, S489F, in the interleukin-31 receptor alpha chain (IL-31Rα). Combined, these results suggest a role for glycosylated hotspot residues in regulating receptor signaling, mutation of which can lead to ligand-independent, uncontrolled activity and human disease.
HostingRepository	PRIDE
AnnounceDate	2023-07-20
AnnouncementXML	Submission_2023-07-20_12:03:08.567.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	MichaelHollander
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	complex glycosylation
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2022-08-01 02:06:13	ID requested
⏵ 1	2023-07-20 12:03:09	announced
2	2023-11-14 09:03:53	announced	2023-11-14: Updated project metadata.

Hollander MJ, Malaker SA, Riley NM, Perez I, Abney NM, Gray MA, Maxson JE, Cochran JR, Bertozzi CR, Mutational screens highlight glycosylation as a modulator of colony-stimulating factor 3 receptor (CSF3R) activity. J Biol Chem, 299(6):104755(2023) [pubmed]

submitter keyword: receptor, mutagenesis, neutrophil,glycoprotein

Carolyn R.Bertozzi
contact affiliation	Department of Chemistry, Stanford University
contact email	bertozzi@stanford.edu
lab head
MichaelHollander
contact affiliation	Stanford University
contact email	hollander@stanford.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD035664
     1. Label: PRIDE project
     2. Name: Mutational screens highlight glycosylation as a modulator of colony-stimulating factor 3 receptor (CSF3R) activity