Ligand-induced EpoR Internalization is Mediated by JAK2 and P85 and is Impaired by Mutations Responsible for Primary Familial and Congenital Polycythemia

Overview

Journal Blood

Publisher Elsevier

Specialty Hematology

Date 2009 Apr 2

PMID 19336760

Citations 35

Authors

Rita Sulahian

Ondine Cleaver

Lily Jun-Shen Huang

Affiliations

Soon will be listed here.

Abstract

Epo-induced endocytosis of EpoR plays important roles in the down-regulation of EpoR signaling and is the primary means that regulates circulating Epo concentrations. Here we show that cell-surface EpoR is internalized via clathrin-mediated endocytosis. Both JAK2 kinase activity and EpoR cytoplasmic tyrosines are important for ligand-dependent EpoR internalization. Phosphorylated Y429, Y431, and Y479 in the EpoR cytoplasmic domain bind p85 subunit of PI3 kinase on Epo stimulation and individually are sufficient to mediate Epo-dependent EpoR internalization. Knockdown of p85alpha and p85beta or expression of their dominant-negative forms, but not inhibition of PI3 kinase activity, dramatically impaired EpoR internalization, indicating that p85alpha and p85beta may recruit proteins in the endocytic machinery on Epo stimulation. Furthermore, mutated EpoRs from primary familial and congenital polycythemia (PFCP) patients lacking the 3 important tyrosines do not bind p85 or internalize on stimulation. Addition of residues encompassing Y429 and Y431 to these truncated receptors restored p85beta binding and Epo sensitivity. Our results identify a novel PI3 kinase activity-independent function of p85 in EpoR internalization and support a model that defects of internalization in truncated EpoRs from PFCP patients contribute to Epo hypersensitivity and prolonged signaling.

Citing Articles

Iron chelation improves ineffective erythropoiesis and iron overload in myelodysplastic syndrome mice.

An W, Feola M, Levy M, Aluri S, Ruiz-Martinez M, Sridharan A Elife. 2023; 12.

PMID: 38153418 PMC: 10754500. DOI: 10.7554/eLife.83103.

Protection of insect neurons by erythropoietin/CRLF3-mediated regulation of pro-apoptotic acetylcholinesterase.

Knorr D, Schneider K, Buschgens L, Forster J, Georges N, Geurten B Sci Rep. 2022; 12(1):18565.

PMID: 36329181 PMC: 9633726. DOI: 10.1038/s41598-022-22035-0.

The Presence of Circulating Nucleated Red Blood Cells Is Associated With Disease Severity in Patients of Hemorrhagic Fever With Renal Syndrome.

Zhang J, Tang K, Zhang Y, Ma Y, Zhang C, Hu H Front Med (Lausanne). 2021; 8:665410.

PMID: 34113638 PMC: 8186265. DOI: 10.3389/fmed.2021.665410.

MEKK3-MEK5-ERK5 signaling promotes mitochondrial degradation.

Craig J, Miller J, Rayavarapu R, Hong Z, Bulut G, Zhuang W Cell Death Discov. 2020; 6:107.

PMID: 33101709 PMC: 7576125. DOI: 10.1038/s41420-020-00342-7.

Erythropoietin regulation of red blood cell production: from bench to bedside and back.

Bhoopalan S, Huang L, Weiss M F1000Res. 2020; 9.

PMID: 32983414 PMC: 7503180. DOI: 10.12688/f1000research.26648.1.

References

Wang J, Auger K, Jarvis L, Shi Y, Roberts T . Direct association of Grb2 with the p85 subunit of phosphatidylinositol 3-kinase. J Biol Chem. 1995; 270(21):12774-80. DOI: 10.1074/jbc.270.21.12774. View

Klingmuller U, Lorenz U, Cantley L, Neel B, Lodish H . Specific recruitment of SH-PTP1 to the erythropoietin receptor causes inactivation of JAK2 and termination of proliferative signals. Cell. 1995; 80(5):729-38. DOI: 10.1016/0092-8674(95)90351-8. View

Beckman D, Lin L, Quinones M, Longmore G . Activation of the erythropoietin receptor is not required for internalization of bound erythropoietin. Blood. 1999; 94(8):2667-75. View

Booker G, Gout I, Downing A, Driscoll P, Boyd J, Waterfield M . Solution structure and ligand-binding site of the SH3 domain of the p85 alpha subunit of phosphatidylinositol 3-kinase. Cell. 1993; 73(4):813-22. DOI: 10.1016/0092-8674(93)90259-s. View

Zhang J, Lodish H . Identification of K-ras as the major regulator for cytokine-dependent Akt activation in erythroid progenitors in vivo. Proc Natl Acad Sci U S A. 2005; 102(41):14605-10. PMC: 1253609. DOI: 10.1073/pnas.0507446102. View

Richmond T, Chohan M, Barber D . Turning cells red: signal transduction mediated by erythropoietin. Trends Cell Biol. 2005; 15(3):146-55. DOI: 10.1016/j.tcb.2005.01.007. View

Sorkin A, Westermark B, Heldin C, Claesson-Welsh L . Effect of receptor kinase inactivation on the rate of internalization and degradation of PDGF and the PDGF beta-receptor. J Cell Biol. 1991; 112(3):469-78. PMC: 2288834. DOI: 10.1083/jcb.112.3.469. View

Sawyer S, HANKINS W . The functional form of the erythropoietin receptor is a 78-kDa protein: correlation with cell surface expression, endocytosis, and phosphorylation. Proc Natl Acad Sci U S A. 1993; 90(14):6849-53. PMC: 47030. DOI: 10.1073/pnas.90.14.6849. View

Huang H, Li L, Wu C, Schibli D, Colwill K, Ma S . Defining the specificity space of the human SRC homology 2 domain. Mol Cell Proteomics. 2007; 7(4):768-84. DOI: 10.1074/mcp.M700312-MCP200. View

10.

Taniguchi C, Tran T, Kondo T, Luo J, Ueki K, Cantley L . Phosphoinositide 3-kinase regulatory subunit p85alpha suppresses insulin action via positive regulation of PTEN. Proc Natl Acad Sci U S A. 2006; 103(32):12093-7. PMC: 1524929. DOI: 10.1073/pnas.0604628103. View

11.

Sorokin A, Mohammadi M, Huang J, Schlessinger J . Internalization of fibroblast growth factor receptor is inhibited by a point mutation at tyrosine 766. J Biol Chem. 1994; 269(25):17056-61. View

12.

Hartley D, Meisner H, Corvera S . Specific association of the beta isoform of the p85 subunit of phosphatidylinositol-3 kinase with the proto-oncogene c-cbl. J Biol Chem. 1995; 270(31):18260-3. DOI: 10.1074/jbc.270.31.18260. View

13.

Glenney Jr J, Chen W, LAZAR C, Walton G, Zokas L, Rosenfeld M . Ligand-induced endocytosis of the EGF receptor is blocked by mutational inactivation and by microinjection of anti-phosphotyrosine antibodies. Cell. 1988; 52(5):675-84. DOI: 10.1016/0092-8674(88)90405-9. View

14.

Gorvel J, Chavrier P, Zerial M, Gruenberg J . rab5 controls early endosome fusion in vitro. Cell. 1991; 64(5):915-25. DOI: 10.1016/0092-8674(91)90316-q. View

15.

Tanaka Y, Tanaka N, Saeki Y, Tanaka K, Murakami M, Hirano T . c-Cbl-dependent monoubiquitination and lysosomal degradation of gp130. Mol Cell Biol. 2008; 28(15):4805-18. PMC: 2493370. DOI: 10.1128/MCB.01784-07. View

16.

Percy M . Genetically heterogeneous origins of idiopathic erythrocytosis. Hematology. 2007; 12(2):131-9. DOI: 10.1080/10245330601111979. View

17.

Holt K, Olson L, Moye-Rowley W, Pessin J . Phosphatidylinositol 3-kinase activation is mediated by high-affinity interactions between distinct domains within the p110 and p85 subunits. Mol Cell Biol. 1994; 14(1):42-9. PMC: 358354. DOI: 10.1128/mcb.14.1.42-49.1994. View

18.

Gout I, Dhand R, Hiles I, Fry M, Panayotou G, Das P . The GTPase dynamin binds to and is activated by a subset of SH3 domains. Cell. 1993; 75(1):25-36. View

19.

Krebs D, Hilton D . SOCS proteins: negative regulators of cytokine signaling. Stem Cells. 2001; 19(5):378-87. DOI: 10.1634/stemcells.19-5-378. View

20.

Jiang X, Sorkin A . Epidermal growth factor receptor internalization through clathrin-coated pits requires Cbl RING finger and proline-rich domains but not receptor polyubiquitylation. Traffic. 2003; 4(8):529-43. DOI: 10.1034/j.1600-0854.2003.t01-1-00109.x. View