{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T12:50:56Z","timestamp":1783687856539,"version":"3.55.0"},"reference-count":50,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,10,29]],"date-time":"2022-10-29T00:00:00Z","timestamp":1667001600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"German Research Foundation (DFG)","award":["Ko1855\/4-1"],"award-info":[{"award-number":["Ko1855\/4-1"]}]},{"name":"German Research Foundation (DFG)","award":["grant 67-0061"],"award-info":[{"award-number":["grant 67-0061"]}]},{"name":"Behring R\u00f6ntgen Foundation","award":["Ko1855\/4-1"],"award-info":[{"award-number":["Ko1855\/4-1"]}]},{"name":"Behring R\u00f6ntgen Foundation","award":["grant 67-0061"],"award-info":[{"award-number":["grant 67-0061"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)","award":["Ko1855\/4-1"],"award-info":[{"award-number":["Ko1855\/4-1"]}]},{"name":"Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)","award":["grant 67-0061"],"award-info":[{"award-number":["grant 67-0061"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Hypoxia stabilizes the transcription factor HIF-1\u03b1, which promotes the transcription of many genes essential to adapt to reduced oxygen levels. Besides proline hydroxylation, expression of HIF-1\u03b1 is also regulated by a range of other posttranslational modifications including phosphorylation by cAMP-dependent protein kinase A (PKA), which stabilizes HIF-1\u03b1. We recently demonstrated that MAGED2 is required for cAMP generation under hypoxia and proposed that this regulation may explain the transient nature of antenatal Bartter syndrome (aBS) due to MAGED2 mutations. Consequently, we sought to determine whether hypoxic induction of HIF-1\u03b1 requires also MAGED2. In HEK293 and HeLa cells, MAGED2 knock-down impaired maximal induction of HIF-1\u03b1 under physical hypoxia as evidenced by time-course experiments, which showed a signification reduction of HIF-1\u03b1 upon MAGED2 depletion. Similarly, using cobalt chloride to induce HIF-1\u03b1, MAGED2 depletion impaired its appropriate induction. Given the known effect of the cAMP\/PKA pathway on the hypoxic induction of HIF-1\u03b1, we sought to rescue impaired HIF-1\u03b1 induction with isoproterenol and forskolin acting upstream and downstream of G\u03b1s, respectively. Importantly, while forskolin induced HIF-1\u03b1 above control levels in MAGED2-depleted cells, isoproterenol had no effect. To further delineate which PKA subtype is involved, we analyzed the effect of two PKA inhibitors and identified that PKA type II regulates HIF-1\u03b1. Interestingly, MAGED2 mRNA and protein were also increased under hypoxia by a cAMP mimetic. Moreover, MAGED2 protein expression also required HIF-1\u03b1. Thus, our data provide evidence for reciprocal regulation of MAGED2 and HIF-1\u03b1 under hypoxia, revealing therefore a new regulatory mechanism that may further explain the transient nature of aBS caused by MAGED2 mutations.<\/jats:p>","DOI":"10.3390\/cells11213424","type":"journal-article","created":{"date-parts":[[2022,10,29]],"date-time":"2022-10-29T23:45:00Z","timestamp":1667087100000},"page":"3424","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Reciprocal Regulation of MAGED2 and HIF-1\u03b1 Augments Their Expression under Hypoxia: Role of cAMP and PKA Type II"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4839-9834","authenticated-orcid":false,"given":"Elie","family":"Seaayfan","sequence":"first","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sadiq","family":"Nasrah","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4400-3129","authenticated-orcid":false,"given":"Lea","family":"Quell","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aline","family":"Radi","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5530-7016","authenticated-orcid":false,"given":"Maja","family":"Kleim","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ralph T.","family":"Schermuly","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Member of the German Center for Lung Research (DZL), Excellence Cluster Cardio-Pulmonary Institute (CPI), Justus-Liebig-University, 35392 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefanie","family":"Weber","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2167-5766","authenticated-orcid":false,"given":"Kamel","family":"Laghmani","sequence":"additional","affiliation":[{"name":"Centre de Recherche des Cordeliers, Sorbonne Universit\u00e9, Inserm, Universit\u00e9 de Paris, CNRS, ERL8228, F-75006 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6499-5313","authenticated-orcid":false,"given":"Martin","family":"K\u00f6mhoff","sequence":"additional","affiliation":[{"name":"University Children\u2019s Hospital, Philipps University, 35043 Marburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.1152\/physrev.00043.2017","article-title":"Gestational Hypoxia and Developmental Plasticity","volume":"98","author":"Ducsay","year":"2018","journal-title":"Physiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1571","DOI":"10.1007\/s00467-016-3333-5","article-title":"Role of hypoxia during nephrogenesis","volume":"31","author":"Hemker","year":"2016","journal-title":"Pediatr. Nephrol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"452","DOI":"10.1203\/00006450-197109000-00003","article-title":"Studies on the Circulation of the Previable Human Fetus","volume":"5","author":"Rudolph","year":"1971","journal-title":"Pediatr. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1038\/sj.ki.5000062","article-title":"Expression of hypoxia-inducible transcription factors in developing human and rat kidneys","volume":"69","author":"Bernhardt","year":"2006","journal-title":"Kidney Int."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"647","DOI":"10.1056\/NEJM199503093321006","article-title":"Hypoxia of the renal medulla\u2014Its implications for disease","volume":"332","author":"Brezis","year":"1995","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1097\/MNH.0000000000000346","article-title":"Pathophysiology of antenatal Bartter\u2019s syndrome","volume":"26","author":"Komhoff","year":"2017","journal-title":"Curr. Opin. Nephrol. Hypertens."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"242","DOI":"10.2215\/CJN.05670517","article-title":"Prevalence of Novel MAGED2 Mutations in Antenatal Bartter Syndrome","volume":"13","author":"Legrand","year":"2018","journal-title":"Clin. J. Am. Soc. Nephrol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1853","DOI":"10.1056\/NEJMoa1507629","article-title":"Polyhydramnios, Transient Antenatal Bartter\u2019s Syndrome, and MAGED2 Mutations","volume":"374","author":"Laghmani","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Seaayfan, E., Nasrah, S., Quell, L., Kleim, M., Weber, S., Meyer, H., Laghmani, K., and K\u00f6mhoff, M. (2022). MAGED2 Is Required under Hypoxia for cAMP Signaling by Inhibiting MDM2-Dependent Endocytosis of G-Alpha-S. Cells, 11.","DOI":"10.3390\/cells11162546"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"16121","DOI":"10.1074\/jbc.REV120.008029","article-title":"Emerging roles of the MAGE protein family in stress response pathways","volume":"295","author":"Gee","year":"2020","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1016\/j.molcel.2010.08.029","article-title":"MAGE-RING protein complexes comprise a family of E3 ubiquitin ligases","volume":"39","author":"Doyle","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1016\/0002-9378(93)90646-Z","article-title":"Effects of long-term, high-altitude hypoxemia on ovine fetal cardiac output and blood flow distribution","volume":"169","author":"Kamitomo","year":"1993","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/j.cell.2012.01.021","article-title":"Hypoxia-inducible factors in physiology and medicine","volume":"148","author":"Semenza","year":"2012","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cellsig.2004.04.010","article-title":"Signalling via the hypoxia-inducible factor-1alpha requires multiple posttranslational modifications","volume":"17","author":"Mazure","year":"2005","journal-title":"Cell Signal."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Albanese, A., Daly, L.A., Mennerich, D., Kietzmann, T., and S\u00e9e, V. (2021). The Role of Hypoxia-Inducible Factor Post-Translational Modifications in Regulating Its Localisation, Stability, and Activity. Int. J. Mol. Sci., 22.","DOI":"10.20944\/preprints202012.0112.v1"},{"key":"ref_16","first-page":"222","article-title":"Effect of beta adrenergic blocking agents on erythropoiesis in rats","volume":"226","author":"Neuwirt","year":"1983","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_17","first-page":"176","article-title":"Effects of beta adrenergic blocking agents on erythropoietin production in rabbits exposed to hypoxia","volume":"193","author":"Fink","year":"1975","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2396","DOI":"10.1016\/j.cellsig.2012.08.007","article-title":"cAMP-dependent protein kinase is essential for hypoxia-mediated epithelial-mesenchymal transition, migration, and invasion in lung cancer cells","volume":"24","author":"Shaikh","year":"2012","journal-title":"Cell Signal."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"10121","DOI":"10.1038\/s41598-017-09549-8","article-title":"Hypoxia induces cancer-associated cAMP\/PKA signalling through HIF-mediated transcriptional control of adenylyl cyclases VI and VII","volume":"7","author":"Simko","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"ra56","DOI":"10.1126\/scisignal.aaf0583","article-title":"Protein kinase A-dependent phosphorylation stimulates the transcriptional activity of hypoxia-inducible factor 1","volume":"9","author":"Bullen","year":"2016","journal-title":"Sci. Signal."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"10879","DOI":"10.1038\/s41598-021-90284-6","article-title":"A reliable set of reference genes to normalize oxygen-dependent cytoglobin gene expression levels in melanoma","volume":"11","author":"Maric","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"556","DOI":"10.1002\/jat.3749","article-title":"The use of cobalt chloride as a chemical hypoxia model","volume":"39","year":"2019","journal-title":"J. Appl. Toxicol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1558","DOI":"10.1016\/j.bbamcr.2007.06.002","article-title":"Intermittent hypoxia changes HIF-1alpha phosphorylation pattern in endothelial cells: Unravelling of a new PKA-dependent regulation of HIF-1alpha","volume":"1773","author":"Toffoli","year":"2007","journal-title":"Biochim. Et Biophys. Acta"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"29083","DOI":"10.1074\/jbc.270.49.29083","article-title":"Hypoxia and mitochondrial inhibitors regulate expression of glucose transporter-1 via distinct Cis-acting sequences","volume":"270","author":"Ebert","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"D92","DOI":"10.1093\/nar\/gku1111","article-title":"The Eukaryotic Promoter Database: Expansion of EPDnew and new promoter analysis tools","volume":"43","author":"Dreos","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"20599","DOI":"10.1074\/jbc.270.35.20599","article-title":"Novel (Rp)-cAMPS Analogs as Tools for Inhibition of cAMP-Kinase in Cell Culture: Basal cAMP-Kinase Activity Modulates Interleukin-1\u03b2 Action","volume":"270","author":"Gjertsen","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3367","DOI":"10.1038\/s41388-020-1223-6","article-title":"Hypoxia and the hypoxia inducible factor 1alpha activate protein kinase A by repressing RII beta subunit transcription","volume":"39","author":"Lucia","year":"2020","journal-title":"Oncogene"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10414","DOI":"10.1073\/pnas.1701782114","article-title":"Mechanisms for restraining cAMP-dependent protein kinase revealed by subunit quantitation and cross-linking approaches","volume":"114","author":"Stengel","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"538","DOI":"10.3389\/fendo.2018.00538","article-title":"The Molecular Basis for Specificity at the Level of the Protein Kinase a Catalytic Subunit","volume":"9","year":"2018","journal-title":"Front. Endocrinol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/0896-6273(89)90116-5","article-title":"Distinct patterns of cAMP-dependent protein kinase gene expression in mouse brain","volume":"3","author":"Cadd","year":"1989","journal-title":"Neuron"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1498","DOI":"10.1093\/ndt\/gfy367","article-title":"MAGE genes in the kidney: Identification of MAGED2 as upregulated during kidney injury and in stressed tubular cells","volume":"34","author":"Cuarental","year":"2019","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Shu, S., Wang, Y., Zheng, M., Liu, Z., Cai, J., Tang, C., and Dong, Z. (2019). Hypoxia and Hypoxia-Inducible Factors in Kidney Injury and Repair. Cells, 8.","DOI":"10.3390\/cells8030207"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1681\/ASN.2006090998","article-title":"Inhibition of hypoxia inducible factor hydroxylases protects against renal ischemia-reperfusion injury","volume":"19","author":"Hill","year":"2008","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1681\/ASN.2006070792","article-title":"Immunohistochemical detection of hypoxia-inducible factor-1alpha in human renal allograft biopsies","volume":"18","author":"Rosenberger","year":"2007","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"42642","DOI":"10.1074\/jbc.M112.399162","article-title":"Multiple evolutionarily conserved Di-leucine like motifs in the carboxyl terminus control the anterograde trafficking of NKCC2","volume":"287","author":"Zaarour","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4487","DOI":"10.1074\/jbc.M115.702514","article-title":"OS9 Protein Interacts with Na-K-2Cl Co-transporter (NKCC2) and Targets Its Immature Form for the Endoplasmic Reticulum-associated Degradation Pathway","volume":"291","author":"Seaayfan","year":"2016","journal-title":"J. Biol. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Bakhos-Douaihy, D., Seaayfan, E., Demaretz, S., Komhoff, M., and Laghmani, K. (2021). Differential Effects of STCH and Stress-Inducible Hsp70 on the Stability and Maturation of NKCC2. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22042207"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Demaretz, S., Seaayfan, E., Bakhos-Douaihy, D., Frachon, N., Komhoff, M., and Laghmani, K. (2021). Golgi Alpha1,2-Mannosidase IA Promotes Efficient Endoplasmic Reticulum-Associated Degradation of NKCC2. Cells, 11.","DOI":"10.3390\/cells11010101"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"947","DOI":"10.1002\/humu.24217","article-title":"New insights into the role of endoplasmic reticulum-associated degradation in Bartter Syndrome Type 1","volume":"42","author":"Shaukat","year":"2021","journal-title":"Hum. Mutat."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Fiszer-Kierzkowska, A., Vydra, N., Wysocka-Wycisk, A., Kronekova, Z., Jarz\u0105b, M., Lisowska, K.M., and Krawczyk, Z. (2011). Liposome-based DNA carriers may induce cellular stress response and change gene expression pattern in transfected cells. BMC Mol. Biol., 12.","DOI":"10.1186\/1471-2199-12-27"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1016\/S1097-2765(00)80251-8","article-title":"Degradation of Proteins from the ER of S. cerevisiae Requires an Intact Unfolded Protein Response Pathway","volume":"5","author":"Casagrande","year":"2000","journal-title":"Mol. Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1245\/ASO.2006.12.011","article-title":"The role of genetic markers--NAP1L1, MAGE-D2, and MTA1--in defining small-intestinal carcinoid neoplasia","volume":"13","author":"Kidd","year":"2006","journal-title":"Ann. Surg. Oncol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2463","DOI":"10.1002\/cam4.1522","article-title":"A novel dual-marker expression panel for easy and accurate risk stratification of patients with gastric cancer","volume":"7","author":"Kanda","year":"2018","journal-title":"Cancer Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1002\/jso.21580","article-title":"Differential gene expression profile of MAGE family in taiwanese patients with colorectal cancer","volume":"102","author":"Chung","year":"2010","journal-title":"J. Surg. Oncol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.lungcan.2006.12.004","article-title":"Differential expression profile of MAGE family in non-small-cell lung cancer","volume":"56","author":"Tsai","year":"2007","journal-title":"Lung Cancer"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1186\/s12943-019-1089-9","article-title":"Role of hypoxia in cancer therapy by regulating the tumor microenvironment","volume":"18","author":"Jing","year":"2019","journal-title":"Mol. Cancer"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3569","DOI":"10.1038\/onc.2012.510","article-title":"The growing complexity of HIF-1alpha\u2019s role in tumorigenesis: DNA repair and beyond","volume":"32","author":"Rohwer","year":"2013","journal-title":"Oncogene"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1038\/nrc3521","article-title":"The emerging mutational landscape of G proteins and G-protein-coupled receptors in cancer","volume":"13","author":"Kufareva","year":"2013","journal-title":"Nat. Rev. Cancer"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1002\/gcc.22854","article-title":"Activating genomic alterations in the Gs alpha gene (GNAS) in 274\u2009694 tumors","volume":"59","author":"Tirosh","year":"2020","journal-title":"Genes Chromosomes Cancer"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e90240","DOI":"10.1172\/jci.insight.90240","article-title":"Hypoxia sensing through beta-adrenergic receptors","volume":"1","author":"Cheong","year":"2016","journal-title":"JCI Insight"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/21\/3424\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:05:31Z","timestamp":1760144731000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/21\/3424"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,29]]},"references-count":50,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["cells11213424"],"URL":"https:\/\/doi.org\/10.3390\/cells11213424","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,29]]}}}