{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T13:39:11Z","timestamp":1793972351896,"version":"4.8.0"},"reference-count":38,"publisher":"American Society of Hematology","issue":"10","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2025,3,6]]},"abstract":"<jats:sec>\n                  <jats:title>Abstract<\/jats:title>\n                  <jats:p>Cullin-5 (Cul5) coordinates the assembly of cullin-RING-E3 ubiquitin ligase complexes that include the suppressors of cytokine signaling (SOCS)-box\u2013containing proteins. The SOCS-box proteins function to recruit specific substrates to the complex for ubiquitination and degradation. In hematopoiesis, SOCS-box proteins are best known for regulating the actions of cytokines that utilize the JAK-STAT signaling pathway. However, the roles of most SOCS-box proteins have not been studied in physiological contexts and any actions for Cul5\/SOCS complexes in signaling by several hematopoietic cytokines, including thrombopoietin (TPO) and interleukin-3 (IL-3), remain unknown. To define additional potential roles for Cul5\/SOCS complexes, we generated mice lacking Cul5 in hematopoiesis; the absence of Cul5 is predicted to impair the SOCS-box\u2013dependent actions of all proteins that contain this motif. Here, we show that Cul5-deficient mice develop excess megakaryopoiesis and thrombocytosis revealing a novel mechanism of negative regulation of megakaryocyte-committed stem cells, a distinct population within the hematopoietic stem cell pool that have been shown to rapidly, perhaps directly, generate megakaryocytes, and which are produced in excess in the absence of Cul5. Cul5-deficient megakaryopoiesis is distinctive in being largely independent of TPO\/myeloproliferative leukemia protein and involves signaling via the \u03b2-common and\/or \u03b2-IL-3 receptors, with evidence of deregulated responses to IL-3. This process is independent of the interferon-\u03b1\/\u03b2 receptor, previously implicated in inflammation-induced activation of stem-like megakaryocyte progenitor cells.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1182\/blood.2024025406","type":"journal-article","created":{"date-parts":[[2025,1,10]],"date-time":"2025-01-10T13:07:07Z","timestamp":1736514427000},"page":"1034-1046","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":8,"title":["Cullin-5 controls the number of megakaryocyte-committed stem cells to prevent thrombocytosis in\u00a0mice"],"prefix":"10.1182","volume":"145","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6022-9910","authenticated-orcid":false,"given":"Maria","family":"Kauppi","sequence":"first","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0079-8986","authenticated-orcid":false,"given":"Craig D.","family":"Hyland","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth M.","family":"Viney","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine A.","family":"White","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8380-5311","authenticated-orcid":false,"given":"Carolyn A.","family":"de Graaf","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6103-340X","authenticated-orcid":false,"given":"AnneMarie E.","family":"Welch","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2364-6299","authenticated-orcid":false,"given":"Jumana","family":"Yousef","sequence":"additional","affiliation":[{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"},{"name":"3Advanced Technology and Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4171-3712","authenticated-orcid":false,"given":"Laura F.","family":"Dagley","sequence":"additional","affiliation":[{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"},{"name":"3Advanced Technology and Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2984-0620","authenticated-orcid":false,"given":"Samantha J.","family":"Emery-Corbin","sequence":"additional","affiliation":[{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"},{"name":"3Advanced Technology and Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ladina","family":"Di Rago","sequence":"additional","affiliation":[{"name":"4Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew J.","family":"Kueh","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"},{"name":"5Olivia Newton-John Cancer Research Institute, Heidelberg, VIC, Australia"},{"name":"6School of Cancer Medicine, La Trobe University, Heidelberg, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7539-7581","authenticated-orcid":false,"given":"Marco J.","family":"Herold","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"},{"name":"5Olivia Newton-John Cancer Research Institute, Heidelberg, VIC, Australia"},{"name":"6School of Cancer Medicine, La Trobe University, Heidelberg, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas J.","family":"Hilton","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey J.","family":"Babon","sequence":"additional","affiliation":[{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"},{"name":"7Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicos A.","family":"Nicola","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0800-8472","authenticated-orcid":false,"given":"Kira","family":"Behrens","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Warren S.","family":"Alexander","sequence":"additional","affiliation":[{"name":"1Blood Cells and Blood Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia"},{"name":"2Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"234","reference":[{"issue":"4","key":"2025032119470567500_bib1","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1002\/iub.1011","article-title":"The SOCS box-adapting proteins for ubiquitination and proteasomal degradation","volume":"64","author":"Linossi","year":"2012","journal-title":"IUBMB Life"},{"issue":"12","key":"2025032119470567500_bib2","doi-asserted-by":"crossref","first-page":"1984","DOI":"10.1002\/pro.3519","article-title":"The molecular details of cytokine signaling via the JAK\/STAT pathway","volume":"27","author":"Morris","year":"2018","journal-title":"Protein Sci"},{"issue":"5","key":"2025032119470567500_bib3","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1016\/S0092-8674(00)80047-1","article-title":"SOCS1 is a critical inhibitor of interferon gamma signaling and prevents the potentially fatal neonatal actions of this cytokine","volume":"98","author":"Alexander","year":"1999","journal-title":"Cell"},{"issue":"4","key":"2025032119470567500_bib4","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1016\/S1074-7613(03)00078-5","article-title":"Suppressor of cytokine signaling-1 is a critical regulator of interleukin-7-dependent CD8+ T cell differentiation","volume":"18","author":"Chong","year":"2003","journal-title":"Immunity"},{"issue":"25","key":"2025032119470567500_bib5","doi-asserted-by":"crossref","first-page":"22755","DOI":"10.1074\/jbc.M303021200","article-title":"Suppressor of cytokine signaling-1 regulates signaling in response to interleukin-2 and other gamma c-dependent cytokines in peripheral T cells","volume":"278","author":"Cornish","year":"2003","journal-title":"J\u00a0Biol Chem"},{"issue":"1","key":"2025032119470567500_bib6","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1038\/ni1287","article-title":"Suppressor of cytokine signaling 1 regulates the immune response to infection by a unique inhibition of type I interferon activity","volume":"7","author":"Fenner","year":"2006","journal-title":"Nat Immunol"},{"issue":"5","key":"2025032119470567500_bib7","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/S1074-7613(01)00132-7","article-title":"SOCS-1\/SSI-1-deficient NKT cells participate in severe hepatitis through dysregulated cross-talk inhibition of IFN-gamma and IL-4 signaling in\u00a0vivo","volume":"14","author":"Naka","year":"2001","journal-title":"Immunity"},{"issue":"6","key":"2025032119470567500_bib8","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1038\/ni931","article-title":"SOCS3 negatively regulates IL-6 signaling in\u00a0vivo","volume":"4","author":"Croker","year":"2003","journal-title":"Nat Immunol"},{"issue":"2","key":"2025032119470567500_bib9","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/S1074-7613(04)00022-6","article-title":"SOCS3 is a critical physiological negative regulator of G-CSF signaling and emergency granulopoiesis","volume":"20","author":"Croker","year":"2004","journal-title":"Immunity"},{"issue":"7","key":"2025032119470567500_bib10","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1038\/nm1071","article-title":"Socs3 deficiency in the brain elevates leptin sensitivity and confers resistance to diet-induced obesity","volume":"10","author":"Mori","year":"2004","journal-title":"Nat Med"},{"issue":"45","key":"2025032119470567500_bib11","doi-asserted-by":"crossref","first-page":"16333","DOI":"10.1073\/pnas.0508023102","article-title":"Genetic reduction of embryonic leukemia-inhibitory factor production rescues placentation in SOCS3-null embryos but does not prevent inflammatory disease","volume":"102","author":"Robb","year":"2005","journal-title":"Proc Natl Acad Sci U\u00a0S\u00a0A"},{"issue":"7","key":"2025032119470567500_bib12","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1038\/ni.3470","article-title":"CIS is a potent checkpoint in NK cell-mediated tumor immunity","volume":"17","author":"Delconte","year":"2016","journal-title":"Nat Immunol"},{"issue":"5","key":"2025032119470567500_bib13","doi-asserted-by":"crossref","first-page":"e20191421","DOI":"10.1084\/jem.20191421","article-title":"NK cell-derived GM-CSF potentiates inflammatory arthritis and is negatively regulated by CIS","volume":"217","author":"Louis","year":"2020","journal-title":"J\u00a0Exp Med"},{"issue":"pt 16","key":"2025032119470567500_bib14","doi-asserted-by":"crossref","first-page":"2813","DOI":"10.1242\/jcs.113.16.2813","article-title":"SOCS: physiological suppressors of cytokine signaling","volume":"113","author":"Krebs","year":"2000","journal-title":"J\u00a0Cell Sci"},{"issue":"4","key":"2025032119470567500_bib15","doi-asserted-by":"crossref","first-page":"207","DOI":"10.3109\/08977194.2012.687375","article-title":"The biology and mechanism of action of suppressor of cytokine signaling 3","volume":"30","author":"Babon","year":"2012","journal-title":"Growth Factors"},{"issue":"1","key":"2025032119470567500_bib16","doi-asserted-by":"crossref","first-page":"1558","DOI":"10.1038\/s41467-018-04013-1","article-title":"The molecular basis of JAK\/STAT inhibition by SOCS1","volume":"9","author":"Liau","year":"2018","journal-title":"Nat Commun"},{"issue":"5","key":"2025032119470567500_bib17","doi-asserted-by":"crossref","first-page":"1466","DOI":"10.1182\/blood-2007-03-079178","article-title":"The SOCS box of suppressor of cytokine signaling-3 contributes to the control of G-CSF responsiveness in\u00a0vivo","volume":"110","author":"Boyle","year":"2007","journal-title":"Blood"},{"issue":"23","key":"2025032119470567500_bib18","doi-asserted-by":"crossref","first-page":"13261","DOI":"10.1073\/pnas.231486498","article-title":"The SOCS box of suppressor of cytokine signaling-1 is important for inhibition of cytokine action in\u00a0vivo","volume":"98","author":"Zhang","year":"2001","journal-title":"Proc Natl Acad Sci U\u00a0S\u00a0A"},{"issue":"7690","key":"2025032119470567500_bib19","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1038\/nature25455","article-title":"Hierarchically related lineage-restricted fates of multipotent haematopoietic stem cells","volume":"554","author":"Carrelha","year":"2018","journal-title":"Nature"},{"key":"2025032119470567500_bib20","doi-asserted-by":"crossref","first-page":"11075","DOI":"10.1038\/ncomms11075","article-title":"Single-cell RNA sequencing reveals molecular and functional platelet bias of aged haematopoietic stem cells","volume":"7","author":"Grover","year":"2016","journal-title":"Nat Commun"},{"issue":"4","key":"2025032119470567500_bib21","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.stem.2015.07.007","article-title":"Inflammation-induced emergency megakaryopoiesis driven by hematopoietic stem cell-like megakaryocyte progenitors","volume":"17","author":"Haas","year":"2015","journal-title":"Cell Stem Cell"},{"issue":"1","key":"2025032119470567500_bib22","doi-asserted-by":"crossref","first-page":"4504","DOI":"10.1038\/s41467-022-31914-z","article-title":"Fate mapping of hematopoietic stem cells reveals two pathways of native thrombopoiesis","volume":"13","author":"Morcos","year":"2022","journal-title":"Nat Commun"},{"issue":"10","key":"2025032119470567500_bib23","doi-asserted-by":"crossref","first-page":"3152","DOI":"10.1002\/stem.2106","article-title":"Brief report: single-cell analysis reveals cell division-independent emergence of megakaryocytes from phenotypic hematopoietic stem cells","volume":"33","author":"Roch","year":"2015","journal-title":"Stem Cells"},{"issue":"7687","key":"2025032119470567500_bib24","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1038\/nature25168","article-title":"Clonal analysis of lineage fate in native haematopoiesis","volume":"553","author":"Rodriguez-Fraticelli","year":"2018","journal-title":"Nature"},{"issue":"1","key":"2025032119470567500_bib25","doi-asserted-by":"crossref","first-page":"111447","DOI":"10.1016\/j.celrep.2022.111447","article-title":"Influenza A virus infection instructs hematopoiesis to megakaryocyte-lineage output","volume":"41","author":"Rommel","year":"2022","journal-title":"Cell Rep"},{"issue":"7470","key":"2025032119470567500_bib26","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1038\/nature12495","article-title":"Platelet-biased stem cells reside at the apex of the haematopoietic stem-cell hierarchy","volume":"502","author":"Sanjuan-Pla","year":"2013","journal-title":"Nature"},{"issue":"1","key":"2025032119470567500_bib27","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1242\/dev.01558","article-title":"Assessing the role of hematopoietic plasticity for endothelial and hepatocyte development by non-invasive lineage tracing","volume":"132","author":"Stadtfeld","year":"2005","journal-title":"Development"},{"issue":"1","key":"2025032119470567500_bib28","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.stem.2013.05.014","article-title":"SLAM family markers resolve functionally distinct subpopulations of hematopoietic stem cells and multipotent progenitors","volume":"13","author":"Oguro","year":"2013","journal-title":"Cell Stem Cell"},{"issue":"1","key":"2025032119470567500_bib29","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.stem.2015.05.003","article-title":"Functionally distinct subsets of lineage-biased multipotent progenitors control blood production in normal and regenerative conditions","volume":"17","author":"Pietras","year":"2015","journal-title":"Cell Stem Cell"},{"key":"2025032119470567500_bib30","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/S0076-6879(10)77007-5","article-title":"A\u00a0practical summary of site-specific recombination, conditional mutagenesis, and tamoxifen induction of CreERT2","volume":"477","author":"Anastassiadis","year":"2010","journal-title":"Methods Enzymol"},{"issue":"10","key":"2025032119470567500_bib31","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1038\/ni1008-1091","article-title":"The Immunological Genome Project: networks of gene expression in immune cells","volume":"9","author":"Heng","year":"2008","journal-title":"Nat Immunol"},{"issue":"3-4","key":"2025032119470567500_bib32","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1080\/08977194.2018.1498487","article-title":"Cytokine control of megakaryopoiesis","volume":"36","author":"Behrens","year":"2018","journal-title":"Growth Factors"},{"issue":"4","key":"2025032119470567500_bib33","doi-asserted-by":"crossref","first-page":"1588","DOI":"10.1182\/blood.V96.4.1588","article-title":"Reassessment of interactions between hematopoietic receptors using common beta-chain and interleukin-3-specific receptor beta-chain-null cells: no evidence of functional interactions with receptors for erythropoietin, granulocyte colony-stimulating factor, or stem cell factor","volume":"96","author":"Scott","year":"2000","journal-title":"Blood"},{"issue":"16","key":"2025032119470567500_bib34","doi-asserted-by":"crossref","first-page":"2139","DOI":"10.1182\/blood.2020005563","article-title":"JAK2-V617F and interferon-\u03b1 induce megakaryocyte-biased stem cells characterized by decreased long-term functionality","volume":"137","author":"Rao","year":"2021","journal-title":"Blood"},{"issue":"10","key":"2025032119470567500_bib35","doi-asserted-by":"crossref","first-page":"e124213","DOI":"10.1172\/jci.insight.124213","article-title":"Aged marrow macrophages expand platelet-biased hematopoietic stem cells via interleukin1B","volume":"5","author":"Frisch","year":"2019","journal-title":"JCI Insight"},{"issue":"3","key":"2025032119470567500_bib36","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1073\/pnas.95.3.1195","article-title":"Hematopoietic stem cell deficiencies in mice lacking c-Mpl, the receptor for thrombopoietin","volume":"95","author":"Kimura","year":"1998","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"6","key":"2025032119470567500_bib37","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1016\/j.stem.2007.10.008","article-title":"Critical role of thrombopoietin in maintaining adult quiescent hematopoietic stem cells","volume":"1","author":"Qian","year":"2007","journal-title":"Cell Stem Cell"},{"issue":"5-6","key":"2025032119470567500_bib38","doi-asserted-by":"crossref","first-page":"651","DOI":"10.3109\/08830189809043013","article-title":"Cytokines in hematopoiesis","volume":"16","author":"Alexander","year":"1998","journal-title":"Int Rev Immunol"}],"container-title":["Blood"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/ashpublications.org\/blood\/article-pdf\/145\/10\/1034\/2357386\/blood_bld-2024-025406-main.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/ashpublications.org\/blood\/article-pdf\/145\/10\/1034\/2357386\/blood_bld-2024-025406-main.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,21]],"date-time":"2025-03-21T20:56:48Z","timestamp":1742590608000},"score":1,"resource":{"primary":{"URL":"https:\/\/ashpublications.org\/blood\/article\/145\/10\/1034\/534839\/Cullin-5-controls-the-number-of-megakaryocyte"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,6]]},"references-count":38,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2025,3,6]]}},"URL":"https:\/\/doi.org\/10.1182\/blood.2024025406","relation":{},"ISSN":["0006-4971","1528-0020"],"issn-type":[{"value":"0006-4971","type":"print"},{"value":"1528-0020","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,6]]}}}