Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells.

Sawyer, Stephen T.; Krantz, Sanford B.; Luna, Juan Abel Nájera

doi:10.1073/pnas.84.11.3690

Cited by 95 publications

(34 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This strongly suggests that association between EpoR and TfR2 is direct. In agreement with this hypothesis, mass spectrometry analysis of EpoR-associated proteins did not reveal 5,7,9, and 12 cultures after CD36 ϩ cell sorting. Bottom panel, expression levels of TfR1 (CD71) and GPA measured by flow cytometric analysis of control and TfR2 knockdown cells on culture day 4.…”

Section: Discussionsupporting

confidence: 73%

“…In particular, chemical cross-linking of radiolabeled Epo to cell surface erythroid cells has frequently revealed the presence of 2 additional proteins with apparent molecular masses of 85 and 100 kDa in the EpoR complex. 2,[5][6][7][8] Unfortunately, because of the low expression level of the EpoR, these proteins could never be identified up to now. The dramatic progress that has been recently introduced by the application of mass spectrometry methods to protein analysis led us to address the question of the identity of the EpoR-associated proteins.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Transferrin receptor 2 is a component of the erythropoietin receptor complex and is required for efficient erythropoiesis

Forejtníková

Vieillevoye

Zermati

et al. 2010

Blood

126

161

View full text Add to dashboard Cite

IntroductionErythropoiesis is mainly regulated by the kidney-produced hormone erythropoietin (Epo), which is absolutely required for the survival and proliferation of erythroid progenitors and their terminal differentiation to red cells. 1 The Epo receptor (EpoR) is a type 1 transmembrane protein that belongs to the class 1 cytokine receptor family. Its expression level in erythroid cells is low whatever the differentiation stage and even the most Epo-sensitive cells, colony-forming units erythroid (CFU-Es), or proerythroblasts express less than 1000 EpoRs per cell at their surface. 2 In the absence of Epo, EpoR is believed to be homodimeric, in which each dimer protein is constitutively associated with a Janus kinase-2 (Jak-2) tyrosine-kinase molecule. Epo binding modifies the organization of the receptor complex leading to Jak-2 activation. Association between Jak-2 and the EpoR occurs during the receptor maturation process, most likely before EpoR leaves the endoplasmic reticulum, and is essential for expression of the receptor at the cell surface. 3 However, the mechanisms that control the maturation and the transport of the EpoR to the cell surface remain largely unknown. Indeed, the number of cell surface EpoRs is not related to the synthesis level of this protein and most of these molecules accumulate in the endoplasmic reticulum in EpoRoverexpressing cells. 4 Both functional and direct evidence also suggests that unidentified proteins are associated with EpoR on the cell surface. In particular, chemical cross-linking of radiolabeled Epo to cell surface erythroid cells has frequently revealed the presence of 2 additional proteins with apparent molecular masses of 85 and 100 kDa in the EpoR complex. 2,5-8 Unfortunately, because of the low expression level of the EpoR, these proteins could never be identified up to now. The dramatic progress that has been recently introduced by the application of mass spectrometry methods to protein analysis led us to address the question of the identity of the EpoR-associated proteins.Here, we have purified the EpoR complex from UT7 erythroleukemic cells that express endogenous EpoRs to identify EpoRassociated proteins. Mass spectrometry analysis of the purified proteins revealed the presence of the type 2 transferrin receptor (TfR2) in the EpoR complex.Mammals express 2 transferrin receptors that share similar overall structures but possess specific functions. Most cells including erythroid precursors internalize iron from plasma diferric transferrin through the ubiquitously expressed transferrin receptor 1 (TfR1). A Tfr1 gene deletion is lethal in mice with severe anemia and is not compensated by TfR2. 9 In contrast, TfR2 expression is tissue-restricted with high expression in the liver 10 where it plays a key role in iron metabolism regulation. Indeed, TfR2 contributes to the adaptation of hepcidin production to the needs of the body by sensing the circulating iron bound to transferrin. 11 Familial inactivating or non-sense mutations in the TfR2 gene are responsible for...

show abstract

Section: Discussionsupporting

confidence: 73%

Section: Introductionmentioning

confidence: 99%

Transferrin receptor 2 is a component of the erythropoietin receptor complex and is required for efficient erythropoiesis

Forejtníková

Vieillevoye

Zermati

et al. 2010

Blood

126

161

View full text Add to dashboard Cite

show abstract

“…The EPO-R is a 507-aminoacid polypeptide that forms homodimers following EPO stimulation (61). The existence of multiple cross-linked complexes (33,34,40,51) and multiple affinities (50,51) suggests the possibility of additional EPO-R subunits. The IL-2-R functions as a multisubunit structure composed of ot, 13, and -y chains (45).…”

mentioning

confidence: 99%

Erythropoietin and interleukin-2 activate distinct JAK kinase family members.

Barber¹,

D’Andrea²

1994

Mol. Cell. Biol.

View full text Add to dashboard Cite

The erythropoietin (EPO) receptor and the interleukin-2 (IL-2) receptor ,-chain subunit are members of the cytokine receptor superfamily. They have conserved primary amino acid sequences in their cytoplasmic domains and activate phosphorylation of common substrates, suggesting common biochemical signaling mechanisms. We have generated a cell line, CTLL- EPO-R, that The erythropoietin (EPO) receptor (EPO-R) and the interleukin-2 (IL-2) receptor 1-chain subunit (IL-2-R1) are related members of the cytokine receptor superfamily and share several extracytoplasmic structural features. In addition, the EPO-R and IL-2-R13 have 45% amino acid identity within the box 1 and box 2 regions of their cytoplasmic regions (9, 14). However, the remaining cytoplasmic regions of the IL-2-R1 and EPO-R are highly divergent. The EPO-R is a 507-aminoacid polypeptide that forms homodimers following EPO stimulation (61). The existence of multiple cross-linked complexes (33,34,40,51) and multiple affinities (50, 51) suggests the possibility of additional EPO-R subunits. The IL-2-R functions as a multisubunit structure composed of ot, 13, and -y chains (45). The IL-2-R-y is a common subunit (IL-2-R-yc) shared with the IL-4 (29, 48) and the IL-7 (28, 44) receptors.Previous studies have suggested that the EPO-R and the IL-2-R activate distinct downstream signals (65), although no studies have investigated the biochemical signaling events in cells that express both receptor types. Activation of the EPO-R (64) or the IL-2-R,B (15) results in the induction of tyrosine kinase activity and tyrosine phosphorylation of several proteins. Although neither receptor has intrinsic tyrosine kinase activity, activation of cytoplasmic tyrosine kinases correlates with mitogenesis for these receptors.Activation of the EPO-R results in the rapid tyrosine phosphorylation of the EPO-R itself (5,8,11,39) 632-2112. Fax: (617) 632-2085. intracellular tyrosine kinases such as shown for the T-cell receptor (42,62).Activation of the IL-2-R results in tyrosine phosphorylation of the IL-2-R,B (4, 15, 36, 52), of the IL-2-R-y (2), and of multiple substrates migrating at 180, 116, 97, 70,55, and 42 kDa (26). The IL-2-R,B associates with various Src family kinases such as Lck in CTLL cells (21,22,37) or Lyn and Fyn in Ba/F3-BO3-IL-2-RP cells (27). However, the IL-2-R,B is tyrosine phosphorylated in cell lines that lack Lck expression, including L cells (3, 30), a cell line of fibroblast origin, suggesting that other cytoplasmic tyrosine kinases may be activated by IL-2. JAK tyrosine kinases are widely expressed in many different tissues and cell lines (12,20,63) and are excellent candidates to be involved in the IL-2 signaling pathway. It is not known which, if any, JAK family kinase members are activated by IL-2.The JAK family of tyrosine kinases plays a role in the signal transduction pathway of cytokine receptors. JAK2 was first identified by PCR amplification of conserved DNA sequences encoding tyrosine kinases domains in hematopoietic cells (20). JAK2 b...

show abstract

“…After binding to specific cell surface receptors, Epo stimulates the proliferation and differentiation of committed erythroid progenitor cells; this activity is associated with the activation of a number of erythroid-specific genes, including those encoding carbonic anhydrase and globin (2,20). Epo receptors have been identified primarily on erythroid precursor cells, such as hematopoietic cell lines (3, 11-14, 26, 35, 41), splenic cells from mice treated with phenylhydrazine (29,35) or infected with Friend virus (21,(35)(36)(37), and fetal liver cells (13,31). More recently, Epo receptors have also been detected on megakaryocytes (11), suggesting that their expression may not be restricted exclusively to cells of the erythroid lineage.…”

mentioning

confidence: 99%

“…Unlike normal erythroid cells, MEL cells do not depend on exogenous Epo for proliferation and show different binding affinities for radioiodinated Epo (25,36,37 (5% C02, 95% air) at 37°C (32). For monolayer cultures, cells were attached to 60-mm petri dishes precoated with fibronectin and induced with dimethyl sulfoxide for erythroid differentiation (32).…”

mentioning

confidence: 99%

Structure and transcription of the mouse erythropoietin receptor gene.

Youssoufian¹,

Zon²,

Orkin³

et al. 1990

Mol. Cell. Biol.

118

View full text Add to dashboard Cite

The complete gene encoding the mouse erythropoietin receptor was isolated by using a cDNA probe derived from a mouse erythroleukemia (MEL) cell library. The gene spans -5 kilobases and is present in a single copy per haploid genome. It contains eight exons, and the nucleotide sequence of the coding region from the genomic DNA is identical to the sequence of one of the MEL cDNA clones except for a single amino acid substitution (Leu-*Val) at codon 163. There is a cluster of three major transcriptional start sites -150 nucleotides upstream of the initiator ATG codon which is conserved in erythropoietin-dependent and -independent erythroleukemic cells, in MEL cells at different stages of differentiation, and in normal bone marrow cells. The promoter region contains a potential binding site for Spl, erythroid-specific transcription factor GF-1, and several CACCC boxes, but not typical TATA or CAAT sequences. A fusion gene containing 452 nucleotides of 5' noncoding sequence linked to a promoterless human growth hormone gene directed the transcription of the latter in MEL cells but not in mouse fibroblasts, T cells, B cells, or macrophagelike cells, suggesting that this promoter functions in an erythroid-specific manner.

show abstract

Identification of the receptor for erythropoietin by cross-linking to Friend virus-infected erythroid cells.

Cited by 95 publications

References 23 publications

Transferrin receptor 2 is a component of the erythropoietin receptor complex and is required for efficient erythropoiesis

Transferrin receptor 2 is a component of the erythropoietin receptor complex and is required for efficient erythropoiesis

Erythropoietin and interleukin-2 activate distinct JAK kinase family members.

Structure and transcription of the mouse erythropoietin receptor gene.

Contact Info

Product

Resources

About