{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T20:32:19Z","timestamp":1789072339135,"version":"build-2803163510"},"reference-count":53,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,5,3]],"date-time":"2026-05-03T00:00:00Z","timestamp":1777766400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["RS-2025-16068581"],"award-info":[{"award-number":["RS-2025-16068581"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"award":["RS-2025-16068581"],"award-info":[{"award-number":["RS-2025-16068581"]}],"id":[{"id":"https:\/\/ror.org\/013aysd81","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Bax, a key member of the B-cell lymphoma 2 (Bcl-2) protein family, is essential for inducing mitochondrial apoptosis. In this study, we employed yeast two-hybrid screening to identify ubiquitin-specific protease 49 (USP49) as a binding partner of Bax. Subsequent immunoprecipitation and glutathione S-transferase (GST) pull-down assays confirmed their direct interaction. Functional assays showed that USP49 reduces Bax polyubiquitination at multiple lysine residues within ubiquitin, with the strongest effects observed on K11, K29, K33, and K63 linkages. In contrast, its effect on K48-linked ubiquitination was weak and insufficient to influence Bax protein stability, indicating that USP49 does not regulate Bax abundance through proteasomal degradation. Instead, RT-qPCR analysis revealed that USP49 overexpression significantly increased Bax mRNA levels, and this effect was maintained under apoptosis stimuli (UV, H2O2, and STS), indicating transcriptional regulation largely independent of stress-induced damage, whereas its effect was modest and not statistically significant under starurosporine treatment. Collectively, these findings demonstrate that USP49 regulates Bax primarily through K29\/K33\/K63-linked ubiquitination and transcriptional upregulation, highlighting its role as a stress-responsive modulator of apoptosis and a potential therapeutic target in cancer. Moreover, under DNA damage condition (UV), USP49 overexpression marked enhanced apoptosis.<\/jats:p>","DOI":"10.3390\/ijms27094102","type":"journal-article","created":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T01:12:09Z","timestamp":1777857129000},"page":"4102","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Ubiquitin-Specific Protease 49 Interacts with Bax to Modulate Apoptosis"],"prefix":"10.3390","volume":"27","author":[{"given":"Hae-Seul","family":"Choi","sequence":"first","affiliation":[{"name":"Department of Bioconvergence, Graduate School, CHA University, Seongnam 13488, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soo-Yeon","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, Graduate School, CHA University, Seongnam 13488, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"So-Ra","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, Graduate School, CHA University, Seongnam 13488, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7662-7190","authenticated-orcid":false,"given":"Kwang-Hyun","family":"Baek","sequence":"additional","affiliation":[{"name":"Department of Bioconvergence, Graduate School, CHA University, Seongnam 13488, Republic of Korea"},{"name":"Department of Biomedical Science, Graduate School, CHA University, Seongnam 13488, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1038\/s41580-023-00629-4","article-title":"Mechanisms of BCL-2 family proteins in mitochondrial apoptosis","volume":"24","author":"Czabotar","year":"2023","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4700","DOI":"10.1038\/s41467-024-49067-6","article-title":"Lipid unsaturation promotes BAX and BAK pore activity during apoptosis","volume":"15","author":"Dadsena","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1007\/s11033-024-09805-x","article-title":"Hepatoprotective potential of taxifolin in type 2 diabetic rats: Modulation of oxidative stress and Bcl2\/Bax\/Caspase-3 signaling pathway","volume":"51","author":"Khadrawy","year":"2024","journal-title":"Mol. Biol. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.bbamcr.2010.12.019","article-title":"Molecular biology of Bax and Bak activation and action","volume":"1813","author":"Westphal","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"a036319","DOI":"10.1101\/cshperspect.a036319","article-title":"BAX, BAK, and BOK: A coming of age for the BCL-2 family effector proteins","volume":"12","author":"Moldoveanu","year":"2020","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1111\/j.1600-0560.2006.00700.x","article-title":"Downregulation of cell cycle modulators p21, p27, p53, Rb and proapoptotic Bcl-2-related proteins Bax and Bak in cutaneous melanoma is associated with worse patient prognosis: Preliminary findings","volume":"34","author":"Tchernev","year":"2007","journal-title":"J. Cutan. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1002\/ijc.10850","article-title":"Multigene analysis of Rb pathway and apoptosis control in esophageal squamous cell carcinoma identifies patients with good prognosis","volume":"103","author":"Guner","year":"2003","journal-title":"Int. J. Cancer"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3850","DOI":"10.1073\/pnas.070047997","article-title":"Bax degradation by the ubiquitin\/proteasome-dependent pathway: Involvement in tumor survival and progression","volume":"97","author":"Li","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1016\/j.ccell.2017.09.001","article-title":"Direct activation of BAX by BTSA1 overcomes apoptosis resistance in acute myeloid leukemia","volume":"32","author":"Reyna","year":"2017","journal-title":"Cancer Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1038\/nrm.2017.83","article-title":"Ubiquitylation at the crossroads of development and disease","volume":"19","author":"Rape","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1038\/s41580-019-0099-1","article-title":"Breaking the chains: Deubiquitylating enzyme specificity begets function","volume":"20","author":"Clague","year":"2019","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1038\/cr.2016.39","article-title":"Ubiquitin modifications","volume":"26","author":"Swatek","year":"2016","journal-title":"Cell Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1016\/j.dnarep.2009.01.013","article-title":"Regulatory ubiquitylation in response to DNA double-strand breaks","volume":"8","author":"Panier","year":"2009","journal-title":"DNA Repair"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.bbamcr.2004.10.003","article-title":"Mechanism and function of deubiquitinating enzymes","volume":"1695","author":"Amerik","year":"2004","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1016\/j.cell.2005.11.007","article-title":"A genomic and functional inventory of deubiquitinating enzymes","volume":"123","author":"Nijman","year":"2005","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1007\/s00018-022-04132-5","article-title":"Pro-apoptotic and anti-apoptotic regulation mediated by deubiquitinating enzymes","volume":"79","author":"Choi","year":"2022","journal-title":"Cell Mol. Life Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1016\/j.molcel.2016.05.009","article-title":"MINDY-1 is a member of an rvolutionarily conserved and structurally distinct new family of deubiquitinating enzymes","volume":"63","author":"Kristariyanto","year":"2016","journal-title":"Mol. Cell"},{"key":"ref_18","first-page":"211","article-title":"Protein ubiquitination: A regulatory post-translational modification","volume":"2","author":"Wilkinson","year":"1987","journal-title":"Anticancer. Drug Des."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1458","DOI":"10.1038\/emboj.2010.39","article-title":"IBRDC2, an IBR-type E3 ubiquitin ligase, is a regulatory factor for Bax and apoptosis activation","volume":"29","author":"Benard","year":"2010","journal-title":"EMBO J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6283","DOI":"10.1073\/pnas.1113248109","article-title":"The ubiquitin E3 ligase parkin regulates the proapoptotic function of Bax","volume":"109","author":"Johnson","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2322","DOI":"10.1038\/s41418-023-01221-1","article-title":"The E3 ubiquitin ligase TRIM17 promotes gastric cancer survival and progression via controlling BAX stability and antagonizing apoptosis","volume":"30","author":"Shen","year":"2023","journal-title":"Cell Death Differ."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2930","DOI":"10.1038\/onc.2016.445","article-title":"EGF-mediated inhibition of ubiquitin-specific peptidase 24 expression has a crucial role in tumorigenesis","volume":"36","author":"Wang","year":"2017","journal-title":"Oncogene"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1265","DOI":"10.1177\/1535370216654587","article-title":"Bax-induced apoptosis shortens the life span of DNA repair defect Ku70-knockout mice by inducing emphysema","volume":"241","author":"Matsuyama","year":"2016","journal-title":"Exp. Biol. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/S1097-2765(04)00094-2","article-title":"Acetylation of the C terminus of Ku70 by CBP and PCAF controls Bax-mediated apoptosis","volume":"13","author":"Cohen","year":"2004","journal-title":"Mol. Cell"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Wang, A., Ning, Z., Lu, C., Gao, W., Liang, J., Yan, Q., Tan, G., and Liu, J. (2017). USP22 Induces Cisplatin Resistance in Lung Adenocarcinoma by Regulating \u03b3H2AX-Mediated DNA Damage Repair and Ku70\/Bax-Mediated Apoptosis. Front. Pharmacol., 8.","DOI":"10.3389\/fphar.2017.00274"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Choi, H.S., Lim, E.S., and Baek, K.H. (2022). Deubiquitinating enzyme USP12 regulates the pro-apoptosis protein Bax. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232113107"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1443","DOI":"10.1016\/j.bbrc.2016.12.055","article-title":"Ubiquitin-specific protease 21 regulating the K48-linked polyubiquitination of NANOG","volume":"482","author":"Kwon","year":"2017","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.15252\/embr.201540891","article-title":"Expanding the ubiquitin code through post-translational modification","volume":"16","author":"Herhaus","year":"2015","journal-title":"EMBO Rep."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1146\/annurev.biochem.67.1.425","article-title":"The ubiquitin system","volume":"67","author":"Hershko","year":"1998","journal-title":"Annu. Rev. Biochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1002\/iub.1402","article-title":"Deubiquitinating enzymes in cellular signaling and disease regulation","volume":"67","author":"Hanpude","year":"2015","journal-title":"IUBMB Life"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e2058","DOI":"10.1038\/cddis.2015.275","article-title":"Potential of apoptotic pathway-targeted cancer therapeutic research: Where do we stand?","volume":"7","author":"Baig","year":"2016","journal-title":"Cell Death Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1039\/c0ib00144a","article-title":"Apoptotic cell signaling in cancer progression and therapy","volume":"3","author":"Plati","year":"2011","journal-title":"Integr. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1111\/j.1442-2042.2005.01051.x","article-title":"Apoptosis in prostate cancer: Bax correlation with stage","volume":"12","author":"Amirghofran","year":"2005","journal-title":"Int. J. Urol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1136\/gut.43.3.414","article-title":"bax, but not bcl-2, influences the prognosis of human pancreatic cancer","volume":"43","author":"Friess","year":"1998","journal-title":"Gut"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"549","DOI":"10.2478\/v10035-011-0087-4","article-title":"Significance of Bax expression in breast cancer patients","volume":"83","author":"Pluta","year":"2011","journal-title":"Pol. Przegl Chir."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2790","DOI":"10.1182\/blood-2007-10-110460","article-title":"Increased proteasomal degradation of Bax is a common feature of poor prognosis chronic lymphocytic leukemia","volume":"111","author":"Agrawal","year":"2008","journal-title":"Blood"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1581","DOI":"10.1101\/gad.211037.112","article-title":"USP49 deubiquitinates histone H2B and regulates cotranscriptional pre-mRNA splicing","volume":"27","author":"Zhang","year":"2013","journal-title":"Genes Dev."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1434","DOI":"10.15252\/embj.201695669","article-title":"USP49 negatively regulates tumorigenesis and chemoresistance through FKBP51-AKT signaling","volume":"36","author":"Luo","year":"2017","journal-title":"EMBO J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e0043421","DOI":"10.1128\/mcb.00434-21","article-title":"USP49-Mediated Histone H2B Deubiquitination Regulates HCT116 Cell Proliferation through MDM2-p53 Axis","volume":"42","author":"Shi","year":"2022","journal-title":"Mol. Cell Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1038\/s41419-018-0475-3","article-title":"USP49 participates in the DNA damage response by forming a positive feedback loop with p53","volume":"9","author":"Tu","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.bbrc.2022.03.138","article-title":"c-MYC-USP49-BAG2 axis promotes proliferation SSand chemoresistance of colorectal cancer cells in vitro","volume":"607","author":"Tu","year":"2022","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"120107","DOI":"10.1016\/j.bbamcr.2026.120107","article-title":"USP49 regulates lipid metabolism in hepatocellular carcinoma by stabilizing RACK1 to promote tumor proliferation and migration","volume":"1873","author":"Xu","year":"2026","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"21085","DOI":"10.1074\/jbc.M400063200","article-title":"Phosphorylation of Bax Ser184 by Akt regulates its activity and apoptosis in neutrophils","volume":"279","author":"Gardai","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"14040","DOI":"10.1074\/jbc.M108975200","article-title":"The phosphatidylinositol 3-kinase (PI3K)-Akt pathway suppresses Bax translocation to mitochondria","volume":"277","author":"Tsuruta","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"7779","DOI":"10.1038\/sj.onc.1204984","article-title":"The protein kinase PKB\/Akt regulates cell survival and apoptosis by inhibiting Bax conformational change","volume":"20","author":"Yamaguchi","year":"2001","journal-title":"Oncogene"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"146599","DOI":"10.1016\/j.gene.2022.146599","article-title":"USP49 is a novel deubiquitylating enzyme for \u03b3 H2AX in DNA double-strand break repair","volume":"833","author":"Matsui","year":"2022","journal-title":"Gene"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2555","DOI":"10.1038\/s41388-022-02267-0","article-title":"USP49 mediates tumor progression and poor prognosis through a YAP1-dependent feedback loop in gastric cancer","volume":"41","author":"Liu","year":"2022","journal-title":"Oncogene"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"bgae060","DOI":"10.1093\/carcin\/bgae060","article-title":"USP49 promotes adenocarcinoma of the esophagogastric junction malignant progression via activating SHCBP1-beta-catenin-GPX4 axis","volume":"46","author":"Ding","year":"2024","journal-title":"Carcinogenesis"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"113802","DOI":"10.1016\/j.intimp.2024.113802","article-title":"Membrane palmitoylated protein MPP1 inhibits immune escape by regulating the USP12\/CCL5 axis in urothelial carcinoma","volume":"146","author":"Wu","year":"2025","journal-title":"Int. Immunopharmacol."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Ye, L., Zhang, Q., Liuyu, T., Xu, Z., Zhang, M.X., Luo, M.H., Zeng, W.B., Zhu, Q., Lin, D., and Zhong, B. (2019). USP49 negatively regulates cellular antiviral responses via deconjugating K63-linked ubiquitination of MITA. PLoS Pathog., 15.","DOI":"10.1371\/journal.ppat.1007680"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1007\/s43188-024-00236-1","article-title":"Lysine-63-linked polyubiquitination: A principal target of cadmium carcinogenesis","volume":"40","author":"Chargui","year":"2024","journal-title":"Toxicol. Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1016\/j.tcb.2011.08.008","article-title":"K11-linked ubiquitin chains as novel regulators of cell division","volume":"21","author":"Wickliffe","year":"2011","journal-title":"Trends Cell Biol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s11658-020-00245-6","article-title":"Beyond K48 and K63: Non-canonical protein ubiquitination","volume":"26","author":"Tracz","year":"2021","journal-title":"Cell Mol. Biol. Lett."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/9\/4102\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T04:28:16Z","timestamp":1778560096000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/9\/4102"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,3]]},"references-count":53,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["ijms27094102"],"URL":"https:\/\/doi.org\/10.3390\/ijms27094102","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,3]]}}}