{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T21:05:17Z","timestamp":1791493517661,"version":"4.3.4"},"reference-count":32,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,1,27]],"date-time":"2022-01-27T00:00:00Z","timestamp":1643241600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31700731"],"award-info":[{"award-number":["31700731"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi Province, China","award":["2019L0007"],"award-info":[{"award-number":["2019L0007"]}]},{"name":"Research Project Supported by Department of Resource and Social Security of Shanxi Province, China","award":["20210034"],"award-info":[{"award-number":["20210034"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The FH gene encodes the fumarate hydratase of the Krebs cycle and functions as a homotetramer to catalyze the hydration of fumarate to malate. Mutations in FH result in uterine leiomyomas, a rare autosomal dominant inherited metabolic disease. However, how FH mutations result in this disease is poorly understood. Here, the FH mutation c.557G&gt;A (p.S186N) was identified in a family with uterine leiomyomas phenotype. A series of studies were performed to confirm the pathogenicity of this mutation. Results showed that the FH mutant exhibited significantly lower fumarase enzyme activity and increased the fumarates level compared with the wildtype, which might be due to the impaired homotetramer formation in the native gel electrophoresis. Interestingly, the immunofluorescence study revealed that the overexpressed FH mutant exhibited puncta structures compared with the evenly expressed FH wildtype in cytoplasm suggesting that the altered amino acid might result in dysfunctional proteins which were accumulated to reduce its cytotoxicity. Importantly, the cells overexpressing the FH mutant exhibited higher proliferation and extracellular acidification rate value (ECAR) which might be caused by the upregulated HIF-1\u03b1 indicating the tumor phenotype. Notably, phospho-mTOR was significantly increased and autophagy was inhibited in the FH mutant overexpression cells compared with the wildtype. Our work provides new insight into the FH mutation c.557G&gt;A (p.S186N) underlies uterine leiomyomas and important information for accurate genetic counseling and clinical diagnosis of the disease.<\/jats:p>","DOI":"10.3390\/ijms23031452","type":"journal-article","created":{"date-parts":[[2022,1,27]],"date-time":"2022-01-27T22:00:56Z","timestamp":1643320856000},"page":"1452","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Functional Characterization of FH Mutation c.557G&gt;A Underlies Uterine Leiomyomas"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6137-6154","authenticated-orcid":false,"given":"Ping","family":"Li","sequence":"first","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanru","family":"Wu","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huizhi","family":"Wu","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiuhong","family":"Xiong","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Na","family":"Zhao","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guangxin","family":"Chen","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changxin","family":"Wu","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1511-9843","authenticated-orcid":false,"given":"Han","family":"Xiao","sequence":"additional","affiliation":[{"name":"Institutes of Biomedical Sciences, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education of China, The Key Laboratory of Medical Molecular Cell Biology of Shanxi Province, Shanxi University, Taiyuan 030006, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.jaad.2017.01.023","article-title":"Hereditary leiomyomatosis and renal cell cancer syndrome: An update and review","volume":"77","author":"Patel","year":"2017","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1086\/376435","article-title":"Mutations in the fumarate hydratase gene cause hereditary leiomyomatosis and renal cell cancer in families in North America","volume":"73","author":"Toro","year":"2003","journal-title":"Am. J. Hum. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1038\/ng849","article-title":"Germline mutations in FH predispose to dominantly inherited uterine fibroids, skin leiomyomata and papillary renal cell cancer","volume":"30","author":"Tomlinson","year":"2002","journal-title":"Nat. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1584","DOI":"10.1001\/archdermatol.2008.517","article-title":"Association of germline mutations in the fumarate hydratase gene and uterine fibroids in women with hereditary leiomyomatosis and renal cell cancer","volume":"144","author":"Stewart","year":"2008","journal-title":"Arch. Dermatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1001\/archderm.141.2.199","article-title":"Clinical features of multiple cutaneous and uterine leiomyomatosis: An underdiagnosed tumor syndrome","volume":"141","author":"Alam","year":"2005","journal-title":"Arch. Dermatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3387","DOI":"10.1073\/pnas.051633798","article-title":"Inherited susceptibility to uterine leiomyomas and renal cell cancer","volume":"98","author":"Launonen","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1503\/cmaj.150191","article-title":"Hereditary leiomyomatosis and renal cell cancer syndrome","volume":"188","author":"Tulandi","year":"2016","journal-title":"CMAJ"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1007\/s10545-011-9294-8","article-title":"Structural basis of fumarate hydratase deficiency","volume":"34","author":"Picaud","year":"2011","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"14518","DOI":"10.3390\/ijms131114518","article-title":"Protein profiling of blood samples from patients with hereditary leiomyomatosis and renal cell cancer by surface-enhanced laser desorption\/ionization time-of-flight mass spectrometry","volume":"13","author":"Kamai","year":"2012","journal-title":"Int. J. Mol. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3844","DOI":"10.1093\/hmg\/ddq305","article-title":"Dysregulation of hypoxia pathways in fumarate hydratase-deficient cells is independent of defective mitochondrial metabolism","volume":"19","author":"Adam","year":"2010","journal-title":"Hum. Mol. Genet."},{"key":"ref_11","first-page":"E7","article-title":"An unusual case of sporadic hereditary leiomyomatosis and renal cell carcinoma syndrome","volume":"95","author":"Nagarajan","year":"2015","journal-title":"Cutis"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Bayley, J.P., Launonen, V., and Tomlinson, I.P. (2008). The FH mutation database: An online database of fumarate hydratase mutations involved in the MCUL (HLRCC) tumor syndrome and congenital fumarase deficiency. BMC Med. Genet., 9.","DOI":"10.1186\/1471-2350-9-20"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2174\/1874091X01812010001","article-title":"Biochemical Characterization of Two Clinically-Relevant Human Fumarase Variants Defective for Oligomerization","volume":"12","author":"Bulku","year":"2018","journal-title":"Open Biochem. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1925","DOI":"10.1111\/febs.14782","article-title":"Structural, biochemical and biophysical characterization of recombinant human fumarate hydratase","volume":"286","author":"Rangel","year":"2019","journal-title":"FEBS J."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4767","DOI":"10.1038\/ncomms5767","article-title":"Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations","volume":"5","author":"Lin","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_16","first-page":"80","article-title":"The Subunits of Fumarase","volume":"17","author":"Hill","year":"1964","journal-title":"Brookhaven Symp. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4207","DOI":"10.1016\/S0021-9258(18)91157-9","article-title":"The Subunits of Fumarase","volume":"239","author":"Kanarek","year":"1964","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3345","DOI":"10.1158\/1078-0432.CCR-13-0304","article-title":"Molecular pathways: Fumarate hydratase-deficient kidney cancer--targeting the Warburg effect in cancer","volume":"19","author":"Linehan","year":"2013","journal-title":"Clin. Cancer Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1016\/j.ctrv.2015.07.002","article-title":"Metabolic alterations in renal cell carcinoma","volume":"41","author":"Massari","year":"2015","journal-title":"Cancer Treat. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2231","DOI":"10.1093\/hmg\/ddi227","article-title":"Accumulation of Krebs cycle intermediates and over-expression of HIF1alpha in tumours which result from germline FH and SDH mutations","volume":"14","author":"Pollard","year":"2005","journal-title":"Hum. Mol. Genet."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.ccr.2005.06.017","article-title":"HIF overexpression correlates with biallelic loss of fumarate hydratase in renal cancer: Novel role of fumarate in regulation of HIF stability","volume":"8","author":"Isaacs","year":"2005","journal-title":"Cancer Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1152\/physiol.00001.2004","article-title":"Hydroxylation of HIF-1: Oxygen sensing at the molecular level","volume":"19","author":"Semenza","year":"2004","journal-title":"Physiology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"10430","DOI":"10.1073\/pnas.190332597","article-title":"Activation of HIF1alpha ubiquitination by a reconstituted von Hippel-Lindau (VHL) tumor suppressor complex","volume":"97","author":"Kamura","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"744","DOI":"10.1038\/s41568-018-0074-8","article-title":"mTOR signalling and cellular metabolism are mutual determinants in cancer","volume":"18","author":"Mossmann","year":"2018","journal-title":"Nat. Rev. Cancer"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7004","DOI":"10.1128\/MCB.22.20.7004-7014.2002","article-title":"Regulation of hypoxia-inducible factor 1alpha expression and function by the mammalian target of rapamycin","volume":"22","author":"Hudson","year":"2002","journal-title":"Mol. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1016\/j.cell.2011.10.026","article-title":"Autophagy: Renovation of cells and tissues","volume":"147","author":"Mizushima","year":"2011","journal-title":"Cell"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1865","DOI":"10.1093\/humrep\/des105","article-title":"Strong family history of uterine leiomyomatosis warrants fumarate hydratase mutation screening","volume":"27","author":"Tolvanen","year":"2012","journal-title":"Hum. Reprod."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1038\/nature10642","article-title":"Reductive carboxylation supports growth in tumour cells with defective mitochondria","volume":"481","author":"Mullen","year":"2011","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/j.ccr.2011.07.018","article-title":"The glycolytic shift in fumarate-hydratase-deficient kidney cancer lowers AMPK levels, increases anabolic propensities and lowers cellular iron levels","volume":"20","author":"Tong","year":"2011","journal-title":"Cancer Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1016\/j.celrep.2018.06.106","article-title":"mTORC1 Upregulation Leads to Accumulation of the Oncometabolite Fumarate in a Mouse Model of Renal Cell Carcinoma","volume":"24","author":"Drusian","year":"2018","journal-title":"Cell Rep."},{"key":"ref_31","first-page":"9874","article-title":"Inner nuclear envelope protein SUN1 plays a prominent role in mammalian mRNA export","volume":"43","author":"Li","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"e864","DOI":"10.1002\/mgg3.864","article-title":"Functional evaluation of a novel GLA causative mutation in Fabry disease","volume":"7","author":"Li","year":"2019","journal-title":"Mol. Genet. Genom. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1452\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:09:08Z","timestamp":1760134148000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1452"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,27]]},"references-count":32,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23031452"],"URL":"https:\/\/doi.org\/10.3390\/ijms23031452","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,27]]}}}