Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis
Abstract Human erythropoiesis is an exquisitely controlled multistep developmental process, and its dysregulation leads to numerous human diseases. Transcriptome and epigenome studies provided insights into system‐wide regulation, but we currently lack a global mechanistic view on the dynamics of pr...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Springer Nature
2020-12-01
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| Series: | Molecular Systems Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.15252/msb.20209813 |
| _version_ | 1797282052538630144 |
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| author | Özge Karayel Peng Xu Isabell Bludau Senthil Velan Bhoopalan Yu Yao Freitas Colaco Ana Rita Alberto Santos Brenda A Schulman Arno F Alpi Mitchell J Weiss Matthias Mann |
| author_facet | Özge Karayel Peng Xu Isabell Bludau Senthil Velan Bhoopalan Yu Yao Freitas Colaco Ana Rita Alberto Santos Brenda A Schulman Arno F Alpi Mitchell J Weiss Matthias Mann |
| author_sort | Özge Karayel |
| collection | DOAJ |
| description | Abstract Human erythropoiesis is an exquisitely controlled multistep developmental process, and its dysregulation leads to numerous human diseases. Transcriptome and epigenome studies provided insights into system‐wide regulation, but we currently lack a global mechanistic view on the dynamics of proteome and post‐translational regulation coordinating erythroid maturation. We established a mass spectrometry (MS)‐based proteomics workflow to quantify and dynamically track 7,400 proteins and 27,000 phosphorylation sites of five distinct maturation stages of in vitro reconstituted erythropoiesis of CD34+ HSPCs. Our data reveal developmental regulation through drastic proteome remodeling across stages of erythroid maturation encompassing most protein classes. This includes various orchestrated changes in solute carriers indicating adjustments to altered metabolic requirements. To define the distinct proteome of each maturation stage, we developed a computational deconvolution approach which revealed stage‐specific marker proteins. The dynamic phosphoproteomes combined with a kinome‐targeted CRISPR/Cas9 screen uncovered coordinated networks of erythropoietic kinases and pinpointed downregulation of c‐Kit/MAPK signaling axis as key driver of maturation. Our system‐wide view establishes the functional dynamic of complex phosphosignaling networks and regulation through proteome remodeling in erythropoiesis. |
| first_indexed | 2024-03-07T17:06:24Z |
| format | Article |
| id | doaj.art-f2d224febe78406ba48cdd980a4aa440 |
| institution | Directory Open Access Journal |
| issn | 1744-4292 |
| language | English |
| last_indexed | 2024-03-07T17:06:24Z |
| publishDate | 2020-12-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj.art-f2d224febe78406ba48cdd980a4aa4402024-03-03T02:40:52ZengSpringer NatureMolecular Systems Biology1744-42922020-12-011612n/an/a10.15252/msb.20209813Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesisÖzge Karayel0Peng Xu1Isabell Bludau2Senthil Velan Bhoopalan3Yu Yao4Freitas Colaco Ana Rita5Alberto Santos6Brenda A Schulman7Arno F Alpi8Mitchell J Weiss9Matthias Mann10Department of Proteomics and Signal Transduction Max Planck Institute of Biochemistry Martinsried GermanyDepartment of Hematology St. Jude Children’s Research Hospital Memphis TN USADepartment of Proteomics and Signal Transduction Max Planck Institute of Biochemistry Martinsried GermanyDepartment of Hematology St. Jude Children’s Research Hospital Memphis TN USADepartment of Hematology St. Jude Children’s Research Hospital Memphis TN USANovo Nordisk Foundation Center for Protein Research Faculty of Health Sciences University of Copenhagen Copenhagen DenmarkNovo Nordisk Foundation Center for Protein Research Faculty of Health Sciences University of Copenhagen Copenhagen DenmarkDepartment of Molecular Machines and Signaling Max Planck Institute of Biochemistry Martinsried GermanyDepartment of Molecular Machines and Signaling Max Planck Institute of Biochemistry Martinsried GermanyDepartment of Hematology St. Jude Children’s Research Hospital Memphis TN USADepartment of Proteomics and Signal Transduction Max Planck Institute of Biochemistry Martinsried GermanyAbstract Human erythropoiesis is an exquisitely controlled multistep developmental process, and its dysregulation leads to numerous human diseases. Transcriptome and epigenome studies provided insights into system‐wide regulation, but we currently lack a global mechanistic view on the dynamics of proteome and post‐translational regulation coordinating erythroid maturation. We established a mass spectrometry (MS)‐based proteomics workflow to quantify and dynamically track 7,400 proteins and 27,000 phosphorylation sites of five distinct maturation stages of in vitro reconstituted erythropoiesis of CD34+ HSPCs. Our data reveal developmental regulation through drastic proteome remodeling across stages of erythroid maturation encompassing most protein classes. This includes various orchestrated changes in solute carriers indicating adjustments to altered metabolic requirements. To define the distinct proteome of each maturation stage, we developed a computational deconvolution approach which revealed stage‐specific marker proteins. The dynamic phosphoproteomes combined with a kinome‐targeted CRISPR/Cas9 screen uncovered coordinated networks of erythropoietic kinases and pinpointed downregulation of c‐Kit/MAPK signaling axis as key driver of maturation. Our system‐wide view establishes the functional dynamic of complex phosphosignaling networks and regulation through proteome remodeling in erythropoiesis.https://doi.org/10.15252/msb.20209813(Phospho)proteomicsCRISPR/Cas9 library screenhuman erythropoiesisSLCsystems biology |
| spellingShingle | Özge Karayel Peng Xu Isabell Bludau Senthil Velan Bhoopalan Yu Yao Freitas Colaco Ana Rita Alberto Santos Brenda A Schulman Arno F Alpi Mitchell J Weiss Matthias Mann Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis Molecular Systems Biology (Phospho)proteomics CRISPR/Cas9 library screen human erythropoiesis SLC systems biology |
| title | Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis |
| title_full | Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis |
| title_fullStr | Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis |
| title_full_unstemmed | Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis |
| title_short | Integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis |
| title_sort | integrative proteomics reveals principles of dynamic phosphosignaling networks in human erythropoiesis |
| topic | (Phospho)proteomics CRISPR/Cas9 library screen human erythropoiesis SLC systems biology |
| url | https://doi.org/10.15252/msb.20209813 |
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