Mutations in the WSAWSE and cytosolic domains of the erythropoietin receptor affect signal transduction and ligand binding and internalization

Dawn E Quelle; Frederick W Quelle; Don M Wojchowski

doi:10.1128/mcb.12.10.4553-4561.1992

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Mutations in the WSAWSE and cytosolic domains of the erythropoietin receptor affect signal transduction and ligand binding and internalization

Journal article

Open access

Peer reviewed

Mutations in the WSAWSE and cytosolic domains of the erythropoietin receptor affect signal transduction and ligand binding and internalization

Dawn E Quelle, Frederick W Quelle and Don M Wojchowski

Molecular and cellular biology, Vol.12(10), pp.4553-4561

10/1992

DOI: 10.1128/mcb.12.10.4553-4561.1992

PMCID: PMC360382

PMID: 1406645

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https://www.ncbi.nlm.nih.gov/pmc/articles/360382View

Open Access

Abstract

The terminal development of erythroid progenitor cells is promoted in part through the interaction of erythropoietin (EPO) with its cell surface receptor. This receptor and a growing family of related cytokine receptors share homologous extracellular features, including a well-conserved WSXWS motif. To explore the functional significance of this motif in the murine EPO receptor, five WSAWSE mutants were prepared and their signal-transducing, ligand binding, and endocytotic properties were compared. EPO receptors mutated at tryptophan residues (W-232, W-235---G; W-235---G; W-235---F) failed to mediate EPO-induced growth or pp100 phosphorylation, while S-236---T and E-237---K mutants exhibited partial to full activity (50 to 100% of wild-type growth and induced phosphorylation). Ligand affinity was reduced for mutant receptors (two- to fivefold), yet expression at the cell surface for all receptors was nearly equivalent. Also, the ability of mutated receptors to internalize ligand was either markedly reduced or abolished (W-235---F), indicating a role for the WSAWSE region in hormone internalization. Interestingly, receptor forms lacking 97% of the cytosolic domain (no signal-transducing capacity; binding affinity reduced two- to threefold) internalized EPO efficiently. This and all WSAWSE receptor forms studied also mediated specific cross-linking of 125I-EPO to three accessory membrane proteins (M(r)s, 120,000, 105,000, and 93,000). These findings suggest that the WSAWSE domain of the EPO receptor is important for EPO-induced signal transduction and ligand internalization. In contrast, although the cytosolic domain is required for growth signaling, it appears nonessential for efficient endocytosis.

Mutagenesis

Ligands

Amino Acid Sequence

Cell Line

Molecular Sequence Data

Glycosylation

Signal Transduction - genetics

Receptors, Erythropoietin - genetics

Animals

Culture Media

Receptors, Erythropoietin - chemistry

Cloning, Molecular

Cytosol - metabolism

Endocytosis - genetics

Mice

Erythropoietin - metabolism

Receptors, Erythropoietin - metabolism

Details

Title: Subtitle: Mutations in the WSAWSE and cytosolic domains of the erythropoietin receptor affect signal transduction and ligand binding and internalization
Creators: Dawn E Quelle - Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802
Frederick W Quelle
Don M Wojchowski
Resource Type: Journal article
Publication Details: Molecular and cellular biology, Vol.12(10), pp.4553-4561
DOI: 10.1128/mcb.12.10.4553-4561.1992
PMID: 1406645
PMCID: PMC360382
NLM abbreviation: Mol Cell Biol
ISSN: 0270-7306
eISSN: 1098-5549
Publisher: United States
Grant note: R29-DK40242 / NIDDK NIH HHS HL44491 / NHLBI NIH HHS
Language: English
Date published: 10/1992
Academic Unit: Pathology; Neuroscience and Pharmacology; Internal Medicine
Record Identifier: 9984040200402771

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