title: Disruption of the mevalonate pathway induces dNTP depletion and DNA damage.
abstract: The mevalonate pathway is tightly linked to cell division. Mevalonate derived non-sterol isoprenoids and cholesterol are essential for cell cycle progression and mitosis completion respectively. In the present work, we studied the effects of fluoromevalonate, a competitive inhibitor of mevalonate diphosphate decarboxylase, on cell proliferation and cell cycle progression in both HL-60 and MOLT-4 cells. This enzyme catalyzes the synthesis of isopentenyl diphosphate, the first isoprenoid in the cholesterol biosynthesis pathway, consuming ATP at the same time. Inhibition of mevalonate diphosphate decarboxylase was followed by a rapid accumulation of mevalonate diphosphate and the reduction of ATP concentrations, while the cell content of cholesterol was barely affected. Strikingly, mevalonate diphosphate decarboxylase inhibition also resulted in the depletion of dNTP pools, which has never been reported before. These effects were accompanied by inhibition of cell proliferation and cell cycle arrest at S phase, together with the appearance of γ-H2AX foci and Chk1 activation. Inhibition of Chk1 in cells treated with fluoromevalonate resulted in premature entry into mitosis and massive cell death, indicating that the inhibition of mevalonate diphosphate decarboxylase triggered a DNA damage response. Notably, the supply of exogenously deoxyribonucleosides abolished γ-H2AX formation and prevented the effects of mevalonate diphosphate decarboxylase inhibition on DNA replication and cell growth. The results indicate that dNTP pool depletion caused by mevalonate diphosphate decarboxylase inhibition hampered DNA replication with subsequent DNA damage, which may have important consequences for replication stress and genomic instability.
authors.affiliation: Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRyCIS, 28034 Madrid, Spain; CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III (ISCIII), Spain. Electronic address: cova67@gmail.com.
authors.forename: Covadonga
authors.initials: C
authors.lastname: Martín Sánchez
authors.affiliation: Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, 28049 Madrid, Spain. Electronic address: jmperezm@gmail.com.
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Pérez Martín
authors.affiliation: Department of Pharmacology, Section of Medical Oncology, Yale School of Medicine, New Haven, CT 06520, USA; Department of Oriental Medicine Resources, College of Environmental & Bioresource Sciences, Chonbuk National University, Jeonju, Jeonbuk, Republic of Korea. Electronic address: jongsik.jin@jbnu.ac.kr.
authors.forename: Jong-Sik
authors.initials: JS
authors.lastname: Jin
authors.affiliation: Laboratory of Functional Foods, IMDEA-Food, 28036 Madrid, Spain. Electronic address: alberto.davalos@imdea.org.
authors.forename: Alberto
authors.initials: A
authors.lastname: Dávalos
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Taipa, Macau, China. Electronic address: wzhang@must.edu.mo.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRyCIS, 28034 Madrid, Spain. Electronic address: gema.delapena@hrc.es.
authors.forename: Gema
authors.initials: G
authors.lastname: de la Peña
authors.affiliation: Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRyCIS, 28034 Madrid, Spain; CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III (ISCIII), Spain. Electronic address: javier.botas@hrc.es.
authors.forename: Javier
authors.initials: J
authors.lastname: Martínez-Botas
authors.affiliation: DNA Replication Group, Spanish National Cancer Research Centre (CNIO), 28029 Madrid, Spain. Electronic address: sarodriguez@cnio.es.
authors.forename: Sara
authors.initials: S
authors.lastname: Rodríguez-Acebes
authors.affiliation: Program in Integrative Cell Signaling and Neurobiology of Metabolism, Section of Comparative Medicine, Department of Pathology and the Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520, USA. Electronic address: yajaira.suarez@yale.edu.
authors.forename: Yajaira
authors.initials: Y
authors.lastname: Suárez
authors.affiliation: Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, 28049 Madrid, Spain. Electronic address: mariajose.hazen@uam.es.
authors.forename: María José
authors.initials: MJ
authors.lastname: Hazen
authors.affiliation: Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRyCIS, 28034 Madrid, Spain; CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III (ISCIII), Spain. Electronic address: diego.gomez@hrc.es.
authors.forename: Diego
authors.initials: D
authors.lastname: Gómez-Coronado
authors.affiliation: Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRyCIS, 28034 Madrid, Spain; CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III (ISCIII), Spain. Electronic address: rebeca.busto@hrc.es.
authors.forename: Rebeca
authors.initials: R
authors.lastname: Busto
authors.affiliation: Department of Pharmacology, Section of Medical Oncology, Yale School of Medicine, New Haven, CT 06520, USA. Electronic address: yccheng@yale.edu.
authors.forename: Yung-Chi
authors.initials: YC
authors.lastname: Cheng
authors.affiliation: Servicio de Bioquímica-Investigación, Hospital Universitario Ramón y Cajal, IRyCIS, 28034 Madrid, Spain; CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III (ISCIII), Spain. Electronic address: miguel.a.lasuncion@hrc.es.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Lasunción
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects; D003853:Deoxyribonucleosides / Q000378:metabolism* / Q000494:pharmacology; D005786:Gene Expression Regulation; D018922:HL-60 Cells; D054879:Halogenation; D045782:Hemiterpenes / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008798:Mevalonic Acid / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D009943:Organophosphorus Compounds / Q000378:metabolism; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D003853:Deoxyribonucleosides; C468783:H2AX protein, human; D045782:Hemiterpenes; D006657:Histones; D009943:Organophosphorus Compounds; D034741:RNA, Small Interfering; C004809:isopentenyl pyrophosphate; C046285:5-diphosphomevalonic acid; D000255:Adenosine Triphosphate; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; D002262:Carboxy-Lyases; C022503:pyrophosphomevalonate decarboxylase; D008798:Mevalonic Acid
keywords: Cholesterol; DNA damage; Deoxyribonucleotides; Fluoromevalonate; Mevalonate