{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:22:27Z","timestamp":1785424947225,"version":"3.56.0"},"reference-count":59,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T00:00:00Z","timestamp":1779753600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016250","name":"New York Medical College","doi-asserted-by":"publisher","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}],"id":[{"id":"10.13039\/100016250","id-type":"DOI","asserted-by":"publisher"}]},{"award":["N\/A"],"award-info":[{"award-number":["N\/A"]}],"id":[{"id":"https:\/\/ror.org\/03dkvy735","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Biomolecular condensates in the cytoplasm and nucleus contribute to carcinogenesis through aberrant signaling by assorted transcription factors and fusion oncoproteins. Oral cancer, which is highly prevalent worldwide, frequently occurs in a U-shaped \u201chigh-risk\u201d zone (floor of mouth, side of tongue, and anterior fauces) which forms the path of liquid transit through the mouth. We previously reported that environmental stresses of saliva-like hypotonicity and beverage-like temperature changes triggered cycles of disassembly\/reassembly of biomolecular condensates of GFP-tagged human myxovirus resistance protein (MxA; alias Mx1) in oral cancer cells. In the present study, we identified some of the constituents of GFP-MxA cytoplasmic condensates in oral cells. These condensates were isolated from interferon (IFN)-\u03bb1-treated GFP-MxA expressing OECM1 human oral cancer cells using magnetic bead-based immunoisolation. Unbiased peptide identification confirmed the presence of MxA\/Mx1 peptides; however, the strongest intensity was for the BACH1 transcription factor family. Immunofluorescence analyses confirmed the association of BACH1 and the family member Nrf2 with cytoplasmic human GFP-MxA condensates. Moreover, GFP-BACH1 and GFP-Nrf2 colocalized with cytoplasmic human HA-MxA condensates in transiently transfected OECM1 cells. Western blot assays confirmed the presence of BACH1 and Nrf2 proteins in complexes isolated using anti-MxA pAb. As much as BACH1 and Nrf2 regulate oxidative stress response genes, it was remarkable that immunofluorescence assays revealed the presence of heme oxygenase 1 (HO1)\u2014a downstream redox regulator\u2014in GFP-MxA condensates. However, these condensates were devoid of p62, KEAP1 and Cul3. In terms of aberrant function, in live cells, the Nrf2 transcription factor underwent rapid disassembly and reassembly cycles driven by saliva-like hypotonicity, and was also disassembled by sulforaphane. The data highlight the unexpected intersections in oral cells between MxA condensates and BACH1, Nrf2 and HO1\u2014proteins well known to be involved in pathways regulating cellular responses to environmental and oxidative stresses, antiviral defense, oral epithelial dysplasia, and cancer progression and metastases.<\/jats:p>","DOI":"10.3390\/cells15110982","type":"journal-article","created":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T16:17:58Z","timestamp":1779812278000},"page":"982","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Pro-Oncogenic Transcription Factors BACH1 and Nrf2 Associate with Cytoplasmic Biomolecular Condensates of GFP-MxA (Myxovirus Resistance Protein A) in Oral Cancer Cells"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9354-9929","authenticated-orcid":false,"given":"Pravin B.","family":"Sehgal","sequence":"first","affiliation":[{"name":"Department of Cell and Molecular Physiology, New York Medical College, Valhalla, NY 10595, USA"},{"name":"Department of Medicine, New York Medical College, Valhalla, NY 10595, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huijuan","family":"Yuan","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Physiology, New York Medical College, Valhalla, NY 10595, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"578","DOI":"10.1002\/1097-0142(196205\/06)15:3<578::AID-CNCR2820150318>3.0.CO;2-3","article-title":"Some observations on oral and pharyngeal carcinomas in the State of Bombay","volume":"15","author":"Paymaster","year":"1962","journal-title":"Cancer"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.ijom.2007.07.028","article-title":"Upregulation of heme oxygenase-1 in oral epithelial dysplasias","volume":"37","author":"Lee","year":"2008","journal-title":"Int. J. Oral Maxillofac. Surg."},{"key":"ref_3","first-page":"11884","article-title":"Essentials of oral cancer","volume":"8","author":"Rivera","year":"2015","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2149","DOI":"10.1002\/1097-0142(197605)37:5<2149::AID-CNCR2820370502>3.0.CO;2-G","article-title":"Anatomical site and size of 222 early asymptomatic oral squamous cell carcinomas","volume":"37","author":"Mashberg","year":"1976","journal-title":"Cancer"},{"key":"ref_5","first-page":"9967097","article-title":"Areca nut chewing: Initiation, addiction, and harmful effects emphasizing the barriers and importance of cessation","volume":"2021","author":"Athukorala","year":"2021","journal-title":"J. Addict."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1056\/NEJMp2203571","article-title":"The seeds of ignorance- consequences of a booming betel-nut economy","volume":"387","author":"Moss","year":"2022","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1351","DOI":"10.1002\/hed.70145","article-title":"Greater Attention to Gingival Cancer and Floor of Mouth Cancer: Based on a Retrospective Analysis of Oral Cancer Across Different Subsites","volume":"48","author":"Li","year":"2026","journal-title":"Head Neck"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e017392","DOI":"10.1136\/bmjgh-2024-017392","article-title":"Association of alcohol and different types of alcoholic beverages on the risk of buccal mucosa cancer in Indian men: A multicentre case-control study","volume":"10","author":"George","year":"2025","journal-title":"BMJ Glob. Health"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Rehman, U., Lakhotia, H., Hao, C.Y., Ahn, Y., Kachole, S., Liu, J., Patel, S., Xu, M.J., Rouhani, M.J., and O\u2019Flynn, P. (2026). Lifetime Prevalence of betel nut chewing in India and Taiwan: Raising awareness of oral cancer risks and the urgent call for regulation. Cancers, 18.","DOI":"10.3390\/cancers18071074"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1002\/ijc.24445","article-title":"High-temperature beverage and foods and esophageal cancer risk\u2014A systematic review","volume":"125","author":"Islami","year":"2009","journal-title":"Int. J. Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"652","DOI":"10.1038\/s41416-025-02953-2","article-title":"Hot beverage intake and esophageal cancer in the UK Biobank: Prospective cohort study","volume":"132","author":"Ramirez","year":"2025","journal-title":"Br. J. Cancer"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/S0022-5320(73)90076-2","article-title":"The permeability of keratinized and non-keratinized oral epithelium to horse radish peroxidase","volume":"43","author":"Squier","year":"1973","journal-title":"J. Ultrastruct. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1177\/00220345890680091101","article-title":"The permeability of human oral mucosa and skin to water","volume":"68","author":"Lesch","year":"1989","journal-title":"J. Dent. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1016\/j.biopha.2016.09.082","article-title":"Role of oral microbiome on oral cancers: A review","volume":"84","author":"Gholizadeh","year":"2016","journal-title":"Biomed. Pharmacother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1166037","DOI":"10.3389\/froh.2023.1166037","article-title":"Influence of site and smoking on malignant transformation in the oral cavity: Is microbiome the missing link?","volume":"4","author":"Galvin","year":"2023","journal-title":"Front. Oral Health"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1038\/s41388-023-02927-9","article-title":"HPV-associated oropharyngeal cancer: In search of surrogate biomarkers for early lesions","volume":"43","author":"Lim","year":"2024","journal-title":"Oncogene"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1038\/nrm.2017.7","article-title":"Biomolecular condensates: Organizers of cellular biochemistry","volume":"18","author":"Banani","year":"2017","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.cell.2018.12.035","article-title":"Considerations and challenges in studying liquid-liquid phase separation and biomolecular condensates","volume":"176","author":"Alberti","year":"2019","journal-title":"Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113691","DOI":"10.1016\/j.ab.2020.113691","article-title":"Biomolecular condensates in cell biology and virology: Phase-separated membraneless organelles (MLOs)","volume":"597","author":"Sehgal","year":"2020","journal-title":"Anal. Biochem."},{"key":"ref_20","first-page":"189245","article-title":"Biomolecular condensates: A new lens on cancer biology","volume":"1880","author":"Jiang","year":"2025","journal-title":"BBA Rev. Cancer"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.ccell.2020.12.003","article-title":"Biomolecular condensates and cancer","volume":"39","author":"Boija","year":"2021","journal-title":"Cancer Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1593","DOI":"10.1016\/j.chembiol.2024.08.009","article-title":"RNA and condensates: Disease implications and therapeutic opportunities","volume":"31","author":"Han","year":"2020","journal-title":"Cell Chem. Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6008","DOI":"10.1038\/s41467-023-41655-2","article-title":"Defining the condensate landscape of fusion oncoproteins","volume":"14","author":"Tripathi","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e01014-19","DOI":"10.1128\/JVI.01014-19","article-title":"Human antiviral protein MxA forms novel metastable membraneless cytoplasmic condensates exhibiting rapid reversible tonicity- driven phase transitions","volume":"93","author":"Davis","year":"2019","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Sehgal, P.B., Yuan, H., and DiSenso-Browne, S.V. (2025). Temperature and WNK-SPAK\/OSR1 kinases dynamically regulate antiviral human GFP-MxA biomolecular condensates in oral cancer cells. Cells, 14.","DOI":"10.20944\/preprints202503.0315.v1"},{"key":"ref_26","first-page":"86","article-title":"Interferon-\u03b1-induced cytoplasmic MxA structures in hepatoma Huh7 and primary endothelial cells","volume":"22","author":"Davis","year":"2018","journal-title":"Contemp. Oncol."},{"key":"ref_27","first-page":"15218","article-title":"Murine GFP-Mx1 forms phase-separated nuclear condensates and associates with cytoplasmic intermediate filaments: Novel antiviral activity against vesicular stomatitis virus","volume":"294","author":"Sehgal","year":"2020","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Sehgal, P.B., Yuan, H., and Jin, Y. (2022). Rapid reversible osmoregulation of cytoplasmic biomolecular condensates of human interferon-\u03b1-induced antiviral MxA GTPase. Int. J. Mol. Sci., 23.","DOI":"10.1101\/2022.08.30.505854"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"946","DOI":"10.1002\/eji.201141657","article-title":"MxA expression induced by \u03b1-defensin in healthy human periodontal tissue","volume":"42","author":"Mahanonda","year":"2012","journal-title":"Eur. J. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"590","DOI":"10.3390\/cells13070590","article-title":"Oral antiviral defesnse: Saliva- and beverage-like hypotonicity dynamically regulate formation of membraneless biomolecular condensates of antiviral human MxA in oral epithelial cells","volume":"13","author":"Sehgal","year":"2024","journal-title":"Cells"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1186\/s40364-024-00570-4","article-title":"Transcription factor BACH1 in cancer: Roles, mechanisms, and prospects for targeted therapy","volume":"12","author":"Hu","year":"2024","journal-title":"Biomark. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"101032","DOI":"10.1016\/j.jbc.2021.101032","article-title":"The transcription factor BACH1 at the crossroads of cancer biology: From epithelial-mesenchymal transition to ferroptosis","volume":"297","author":"Igarashi","year":"2021","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Padilla, J., and Lee, J. (2021). A novel therapeutic target, BACH1, regulates cancer metabolism. Cells, 10.","DOI":"10.3390\/cells10030634"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4888","DOI":"10.7150\/ijbs.98846","article-title":"Antioxidant genes in cancer and metabolic diseases: Focusing on Nrf2, sestrin, and heme oxygenase 1","volume":"20","author":"Shrestha","year":"2024","journal-title":"Int. J. Biol. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.canlet.2019.01.003","article-title":"BTB and CNC homology 1 (Bach1) promotes human ovarian cancer cell metastasis by HMGA2-mediated epithelial-mesenchymal transition","volume":"445","author":"Han","year":"2019","journal-title":"Cancer Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1016\/j.cell.2019.06.003","article-title":"Nrf2 activation promotes lung cancer metastasis by inhibiting the degradation of Bach1","volume":"178","author":"Lignitto","year":"2019","journal-title":"Cell"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Herengt, A., Thyrsted, J., and Holm, C.K. (2012). NRF2 in viral infection. Antioxidants, 10.","DOI":"10.3390\/antiox10091491"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"104135","DOI":"10.1016\/j.redox.2026.104135","article-title":"NRF2 controls a diverse network of antiviral effectors with p62 acting as a central restriction factor effective across virus families","volume":"93","author":"Pedersen","year":"2026","journal-title":"Redox Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.ajpath.2016.11.011","article-title":"Modulation of antiviral immunity by heme oxygenase 1","volume":"187","author":"Espinosa","year":"2017","journal-title":"Am. J. Pathol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1016\/j.freeradbiomed.2011.04.027","article-title":"Nrf2 expression modifies influenza A entry and replication in nasal epithelial cells","volume":"51","author":"Kesic","year":"2011","journal-title":"Free Radic. Biol. Med."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1186\/1465-9921-13-43","article-title":"Nrf2 protects human alveolar epithelial cells against injury induced by influenza A virus","volume":"13","author":"Kosmider","year":"2012","journal-title":"Respir. Res."},{"key":"ref_42","first-page":"102247","article-title":"Regulation of innate immunity by Nrf2","volume":"78","author":"Laguette","year":"2023","journal-title":"Curr. Opin. Immunol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"29893","DOI":"10.1074\/jbc.M115.681494","article-title":"Oligomerization and GTP-binding requirements of MxA for viral target recognition and antiviral activity against influenza A virus","volume":"290","author":"Nigg","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.cell.2015.12.038","article-title":"ATPase-modulated stress granules contain a diverse proteome and substructure","volume":"164","author":"Jain","year":"2016","journal-title":"Cell"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1002\/iub.2689","article-title":"p62 bodies: Phase separation, NRF2 activation, and selective autophagic degradation","volume":"74","author":"Komatsu","year":"2022","journal-title":"IUBMB Life"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e00644-17","DOI":"10.1128\/MCB.00644-17","article-title":"p62-dependent phase separation of patient-derived KEAP1 mutations and NRF2","volume":"38","author":"Cloer","year":"2018","journal-title":"Mol. Cell. Biol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1186\/s13578-023-00978-9","article-title":"Enhanced liquidity of p62 droplets mediated by Smurf1 links Nrf2 activation and autophagy","volume":"13","author":"Xia","year":"2023","journal-title":"Cell Biosci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1034\/j.1600-0854.2002.31003.x","article-title":"Interferon-induced mx proteins: Dynamin-like GTPases with antiviral activity","volume":"3","author":"Haller","year":"2002","journal-title":"Traffic"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1016\/j.biochi.2007.04.015","article-title":"Interferon-induced Mx proteins in antiviral host defense","volume":"89","author":"Haller","year":"2007","journal-title":"Biochimie"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.tim.2014.12.003","article-title":"Mx GTPases: Dynamin-like antiviral machines of innate immunity","volume":"23","author":"Haller","year":"2015","journal-title":"Trends Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1128\/MMBR.00024-13","article-title":"Mx proteins: Antiviral gatekeepers that restrain the uninvited","volume":"77","author":"Verhelst","year":"2013","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"4498","DOI":"10.1128\/jvi.65.8.4498-4501.1991","article-title":"Inhibition of vesicular stomatitis virus mRNA synthesis by human MxA protein","volume":"65","author":"Staeheli","year":"1991","journal-title":"J. Virol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1016\/S0042-6822(95)80071-9","article-title":"Vesicular stomatitis virus transcription inhibited by purified MxA protein","volume":"206","author":"Schwemmle","year":"1996","journal-title":"Virology"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e00125","DOI":"10.1128\/JVI.00125-20","article-title":"Subcellular localization of MxB determines its antiviral potential against influenza virus","volume":"94","author":"Steiner","year":"2020","journal-title":"J. Virol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"8202","DOI":"10.1128\/JVI.74.17.8202-8206.2000","article-title":"A monomeric GTPase-negative MxA mutant with antiviral activity","volume":"74","author":"Janzen","year":"2000","journal-title":"J. Virol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"7278","DOI":"10.1074\/jbc.M004227200","article-title":"Transcription factor BACH1 is recruited to the nucleus by its novel alternative spliced isoform","volume":"276","author":"Kanezaki","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1597","DOI":"10.4103\/jcrt.JCRT_282_19","article-title":"MALAT1 and BACH1 are prognostic biomarkers for triple negative breast cancer","volume":"15","author":"Ou","year":"2019","journal-title":"J. Cancer Res. Ther."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"2776","DOI":"10.1158\/0008-5472.CAN-23-1167","article-title":"WNK1 interaction with KEAP1 promotes NRF2 stabilization to enhance the oxidative stress response in hepatocellular carcinoma","volume":"84","author":"Li","year":"2024","journal-title":"Cancer Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"e202411133","DOI":"10.1083\/jcb.202411133","article-title":"Disruption of macrophage volume drives inflammatory responses and type I interferon signaling","volume":"225","author":"Cook","year":"2026","journal-title":"J. Cell Biol."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/11\/982\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T04:17:52Z","timestamp":1780460272000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/11\/982"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,26]]},"references-count":59,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["cells15110982"],"URL":"https:\/\/doi.org\/10.3390\/cells15110982","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,26]]}}}