title: Efficient use of exogenous isoprenols for protein isoprenylation by MDA-MB-231 cells is regulated independently of the mevalonate pathway.
abstract: Mammalian cells can use exogenous isoprenols to generate isoprenoid diphosphate substrates for protein isoprenylation, but the mechanism, efficiency, and biological importance of this process are not known. We developed mass spectrometry-based methods using chemical probes and newly synthesized stable isotope-labeled tracers to quantitate incorporation of exogenously provided farnesol, geranylgeraniol, and unnatural analogs of these isoprenols containing an aniline group into isoprenoid diphosphates and protein isoprenylcysteines by cultured human cancer cell lines. We found that at exogenous isoprenol concentrations >10 μM, this process can generate as much as 50% of the cellular isoprenoid diphosphate pool used for protein isoprenylation. Mutational activation of p53 in MDA-MB-231 breast cancer cells up-regulates the mevalonate pathway to promote tumor invasiveness. p53 silencing or pharmacological inhibition of HMG-CoA reductase in these cells decreases protein isoprenylation from endogenously synthesized isoprenoids but enhances the use of exogenous isoprenols for this purpose, indicating that this latter process is regulated independently of the mevalonate pathway. Our observations suggest unique opportunities for design of cancer cell-directed therapies and may provide insights into mechanisms underlying pleiotropic therapeutic benefits and unwanted side effects of mevalonate pathway inhibition.
authors.affiliation: Department of Molecular and Cellular Biochemistry, Markey Cancer Center, Kentucky Center for Structural Biology, and Department of Chemistry.
authors.forename: Fredrick
authors.initials: F
authors.lastname: Onono
authors.affiliation: Department of Molecular and Cellular Biochemistry, Markey Cancer Center, Kentucky Center for Structural Biology, and Department of Chemistry.
authors.forename: Thangaiah
authors.initials: T
authors.lastname: Subramanian
authors.affiliation: Division of Cardiovascular Medicine, University of Kentucky College of Medicine, Lexington, Kentucky 40536-0200.
authors.forename: Manjula
authors.initials: M
authors.lastname: Sunkara
authors.affiliation: Division of Cardiovascular Medicine, University of Kentucky College of Medicine, Lexington, Kentucky 40536-0200.
authors.forename: Karunai Leela
authors.initials: KL
authors.lastname: Subramanian
authors.affiliation: Department of Molecular and Cellular Biochemistry, Markey Cancer Center, Kentucky Center for Structural Biology, and Department of Chemistry. Electronic address: hps@uky.edu.
authors.forename: H Peter
authors.initials: HP
authors.lastname: Spielmann
authors.affiliation: Division of Cardiovascular Medicine, University of Kentucky College of Medicine, Lexington, Kentucky 40536-0200; Lexington Veterans Affairs Medical Center, Lexington, Kentucky 40502. Electronic address: a.j.morris@uky.edu.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Morris
mesh_terms: D000970:Antineoplastic Agents / Q000493:pharmacokinetics / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004224:Diterpenes / Q000493:pharmacokinetics / Q000494:pharmacology*; D005204:Farnesol / Q000031:analogs & derivatives / Q000493:pharmacokinetics / Q000494:pharmacology*; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000235:genetics / Q000378:metabolism; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000493:pharmacokinetics / Q000494:pharmacology*; D008798:Mevalonic Acid / Q000378:metabolism*; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D017368:Protein Prenylation / Q000187:drug effects / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D004224:Diterpenes; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D011106:Polyisoprenyl Phosphates; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D005204:Farnesol; C017338:geranylgeraniol; D006903:Hydroxymethylglutaryl CoA Reductases; D008798:Mevalonic Acid
keywords: Breast Cancer; Cholesterol; Isoprenoid; Mass Spectrometry (MS); Protein Isoprenylation
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