Abstract
Compounds that interfere with the synthesis of either mitochondrial DNA or mtDNA-encoded proteins reduce the levels of 13 proteins essential for oxidative phosphorylation, leading to a decrease in mitochondrial adenosine triphosphate (ATP) production. Toxicity caused by these compounds is seldom identified in 24- to 72-h cytotoxicity assays due to the low turnover rates of both mtDNA and mtDNA-encoded proteins. To address this problem, the authors developed a 96-well format, high-content screening (HCS) assay that measures, in eukaryotic cells, the level of Complex IV-subunit 1, an mtDNA-encoded protein synthesized on mitochondrial ribosomes, and the level of Complex V-alpha subunit, a nuclear DNA-encoded protein synthesized on cytosolic ribosomes. The effect of several antibiotics and antiretrovirals on these 2 proteins was assessed, in transformed human liver epithelial cells, 6 days after compound treatment. The results confirmed effects of drugs known to reduce mtDNA-encoded protein levels and also revealed novel information showing that several fluoroquinolones and a macrolide, josamycin, impaired expression of mtDNA-encoded proteins. The HCS assay was robust with an average Z' factor of 0.62. The assay enables large-scale...Continue Reading
References
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Nov 25, 2000·Journal of Molecular Biology·S M PoulsenB Vester
Feb 19, 2002·Antimicrobial Agents and Chemotherapy·Gabriel BirkusTomas Cihlar
Aug 2, 2002·Molecular Cell·Jeffrey L HansenThomas A Steitz
Oct 5, 2002·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Bonnie J HansonSteven W Sherwood
Jul 16, 2003·Journal of Molecular Biology·Tanel TensonMåns Ehrenberg
Oct 4, 2003·Nature Reviews. Drug Discovery·William LewisWilliam C Copeland
Aug 3, 2004·Nature Reviews. Drug Discovery·Ismail Kola, John Landis
Aug 30, 2005·Antimicrobial Agents and Chemotherapy·Eva E NagiecKaren L Leach
May 26, 2006·Antimicrobial Agents and Chemotherapy·E E McKeeT A Marks
Dec 13, 2006·Biochimie·Hongliang ZhangYves Pommier
Sep 11, 2007·Drug Discovery Today·James A Dykens, Yvonne Will
May 27, 2008·Trends in Pharmacological Sciences·Kendall B Wallace
Jan 21, 2009·Journal of Immunological Methods·Sashi NadanacivaYvonne Will
Dec 17, 2009·The Journal of Biological Chemistry·Yiying ZhangZhongcong Xie
Citations
Dec 15, 2011·Toxicological Sciences : an Official Journal of the Society of Toxicology·Zhiwu Lin, Yvonne Will
Jul 16, 2014·Expert Opinion on Drug Metabolism & Toxicology·Yvonne Will, James Dykens
Feb 11, 2015·The International Journal of Biochemistry & Cell Biology·Eligio F IannettiWerner J H Koopman
Sep 23, 2014·Regulatory Toxicology and Pharmacology : RTP·Christine M HuntAyako Suzuki
Apr 25, 2013·Journal of Medicinal Chemistry·Tatiana StoilovaMario Salmona
Apr 9, 2014·Journal of Biomolecular Screening·Shantanu SinghAuguste Genovesio
Dec 8, 2017·Frontiers in Molecular Biosciences·Matthew J Young
May 10, 2011·Current Protocols in Toxicology·Sashi NadanacivaYvonne Will
Mar 23, 2018·Nature·Jo Marchant
Jan 10, 2020·The Journal of Biological Chemistry·Amita GuptaChaitan Khosla
Sep 1, 2018·Nucleic Acids Research·Anu HangasSteffi Goffart
Apr 28, 2019·International Journal of Molecular Sciences·Steffi GoffartJaakko L O Pohjoismäki
May 15, 2019·Biology·Yvonne WillKendall B Wallace
Jul 30, 2020·BMJ Open Ophthalmology·Nasim SalimiaghdamM Cristina Kenney
Dec 19, 2020·The Journal of Biological Chemistry·Carolyn K J YoungMatthew J Young
Jun 28, 2021·Ecotoxicology and Environmental Safety·Ningjuan LiangFang Xiao
Aug 29, 2019·ACS Omega·Cole A FiefJoseph E Deweese