Skip to main content
SpringerLink
Log in

Tetracaine, a local anesthetic, preferentially induces translational inhibition with processing body formation rather than phosphorylation of eIF2α in yeast

  • Research Article
  • Published:
Current Genetics Aims and scope Submit manuscript

Abstract

It is unclear whether local anesthetics, such as tetracaine, and antipsychotics, such as phenothiazines, act on lipids or proteins. In Saccharomyces cerevisiae, these drugs inhibit growth, translation initiation, and actin polarization, and induce cell lysis at high concentrations. These activities are likely due to the cationic amphiphilic structure common to these agents. Although drug-induced translational inhibition is conserved in mammalian cells, other mechanisms, including the phosphorylation of eIF2α, a eukaryotic translational initiation factor, remain poorly understood. At a concentration of 10 mM, tetracaine rapidly inhibited translation initiation and lysed cells, whereas, at 2.5 mM, it slowly induced inhibition without lysis. The pat1 disruptant defective in mRNA decapping and the xrn1 disruptant defective in 5′–3′ exoribonuclease were partially resistant to translational inhibition by tetracaine at each concentration, but the gcn2 disruptant defective in the eIF2α kinase was not. Phosphorylation of eIF2α was induced by 10 mM but not by 2.5 mM tetracaine, whereas processing bodies (P-bodies) were formed at 2.5 mM in Pat1-dependent and -independent manners. Therefore, administration of tetracaine inhibits translation initiation with P-body formation at both concentrations but acts via the Gcn2-eIF2α system only at the higher concentration. Because other local anesthetics and phenothiazines induced Pat1-dependent P-body formation, the mechanisms involved in translational inhibition by these cationic amphiphiles are similar. These results suggest that this dose-dependent biphasic translational inhibition by tetracaine results from an increase in membrane proteins that are indirectly inhibited by nonspecific interactions of cationic amphiphiles with membrane lipids.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Banerjee D, Redman CM (1977) Effect of local anesthetics on plasma protein secretion by rat hepatocytes. Biochim Biophys Acta 500:49–60

    Article  CAS  PubMed  Google Scholar 

  • Bruce DL (1975) Halothane inhibition of RNA and protein synthesis of PHA-treated human lymphocytes. Anesthesiology 42:11–14

    Article  CAS  PubMed  Google Scholar 

  • Butterworth JF, Strichartz GR (1990) Molecular mechanisms of local anesthesia. Anesthesiology 72:711–734

    Article  CAS  PubMed  Google Scholar 

  • Coller J, Parker R (2005) General translational repression by activators of mRNA decapping. Cell 122:875–886

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Curtis BM, Scurlock JE (1981) The mechanism of action of local anesthesia by tetraethylammonium derivatives. Anesthesiology 54:270–277

    Article  CAS  PubMed  Google Scholar 

  • De Filippi L, Fournier M, Cameroni E, Linder P, De Virgilio C, Foti M, Deloche O (2007) Membrane stress is coupled to a rapid translational control of gene expression in chlorpromazine-treated cells. Curr Genet 52:171–185

    Article  CAS  PubMed  Google Scholar 

  • Decker CJ, Parker R (2012) P-bodies and stress granules: possible roles in the control of translation and mRNA degradation. Cold Spring Harb Perspect Biol 4. doi:10.1101/cshperspect.a012286

  • Delley PA, Hall MN (1999) Cell wall stress depolarizes cell growth via hyperactivation of RHO1. J Cell Biol 147:163–174

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Deloche O, de la Cruz J, Kressler D, Doère M, Linder P (2004) A membrane transport defect leads to a rapid attenuation of translation initiation in Saccharomyces cerevisiae. Mol Cell 13:357–366

    Article  CAS  PubMed  Google Scholar 

  • Franks NP, Lieb WR (1990) Mechanisms of general anesthesia. Environ Health Perspect 87:199–205

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ghantous HN, Fernando J, Gandolfi AJ, Brendel K (1991) Minimal biotransformation and toxicity of desflurane in guinea pig liver slices. Anesth Analg 72:796–800

    Article  CAS  PubMed  Google Scholar 

  • Gorga FR, Lienhard GE (1981) Equilibria and kinetics of ligand binding to the human erythrocyte glucose transporter. Evidence for an alternating conformation model for transport. Biochemistry 20:5108–5113

    Article  CAS  PubMed  Google Scholar 

  • Guthrie C, Fink GR (1991) Guide to yeast genetics and molecular biology, vol 194. Academic, USA

    Book  Google Scholar 

  • Heys SD, Norton AC, Dundas CR, Eremin O, Ferguson K, Garlick PJ (1989) Anaesthetic agents and their effect on tissue protein synthesis in the rat. Clin Sci 77:651–655

    CAS  PubMed  Google Scholar 

  • Hinnebusch AG (2005) Translational regulation of GCN4 and the general amino acid control of yeast. Annu Rev Microbiol 59:407–450

    Article  CAS  PubMed  Google Scholar 

  • Holmes LE, Campbell SG, De Long SK, Sachs AB, Ashe MP (2004) Loss of translational control in yeast compromised for the major mRNA decay pathway. Mol Cell Biol 24:2998–3010

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hong DY, Kwon K, Lee KR, Choi YJ, Goo TW, Yu K, Kim SW, Kwon OY (2011) Lidocaine induces endoplasmic reticulum stress-associated apoptosis in vitro and in vivo. Int J Mol Sci 12:7652–7661

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Horber FF, Krayer S, Rehder K, Haymond MW (1988) Anesthesia with halothane and nitrous oxide alters protein and amino acid metabolism in dogs. Anesthesiology 69:319–326

    Article  CAS  PubMed  Google Scholar 

  • Hoyle NP, Castelli LM, Campbell SG, Holmes LEA, Ashe MP (2007) Stress-dependent relocalization of translationally primed mRNPs to cytoplasmic granules that are kinetically and spatially distinct from P-bodies. J Cell Biol 179:65–74

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Keil RL, Wolfe D, Reiner T, Peterson CJ, Riley JL (1996) Molecular genetic analysis of volatile-anesthetic action. Mol Cell Biol 16:3446–3453

    CAS  PubMed Central  PubMed  Google Scholar 

  • Kilchert C, Weidner J, Prescianotto-Baschong C, Spang A (2010) Defects in the secretory pathway and high Ca2 induce multiple P-bodies. Mol Biol Cell 21:2624–2638

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kumar RV, Panniers R, Wolfman A, Henshaw EC (1991) Inhibition of protein synthesis by antagonists of calmodulin in Ehrlich ascites tumor cells. Eur J Biochem 195:313–319

    Article  CAS  PubMed  Google Scholar 

  • Kushnirov VV (2000) Rapid and reliable protein extraction from yeast. Yeast 16:857–860

    Article  CAS  PubMed  Google Scholar 

  • Lee IS, Park TJ, Suh BC, Kim YS, Rhee IJ, Kim KT (1999) Chlorpromazine-induced inhibition of catecholamine secretion by a dinicotinic receptors and L-type Ca2 channels in rat pheochromocytoma cells. Biochem Pharmacol 58:1017–1024

    Article  CAS  PubMed  Google Scholar 

  • Meyer H (1899) Theorie der Alkoholnarkose. Arch Exp Pathol Pharmakol 42:109–118

    Article  Google Scholar 

  • Mia M (1975) The effect of psychotropic substances (aminazin, majeptil, trisedil) on protein synthesis in different regions of the rat brain. Biull Eksp Biol Med 79:62–64

    Google Scholar 

  • Mozrzymas JW, Barberis A, Michalak K, Cherubini E (1999) Chlorpromazine inhibits miniature GABAergic currents by reducing the binding and by increasing the unbinding rate of GABAA receptors. J Neurosci 19:2474–2488

    CAS  PubMed  Google Scholar 

  • Nissan T, Rajyaguru P, She M, Song H, Parker R (2010) Decapping activators in Saccharomyces cerevisiae act by multiple mechanisms. Mol Cell 39:773–783

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ogata N, Yoshii M, Narahashi T (1989) Psychotropic drugs block voltage-gated ion channels in neuroblstoma cells. Brain Res 474:140–144

    Article  Google Scholar 

  • Oka Y, Asano T, Shibasaki Y, Lin JL, Tsukuda K, Katagiri H, Akanuma Y, Takaku F (1990) C-terminal truncated glucose transporter is locked into an inward-facing form without transport activity. Nature 345:550–553

    Article  CAS  PubMed  Google Scholar 

  • Ozcan S, Johnston M (1999) Function and regulation of yeast hexose transporters. Microbiol Mol Biol Rev 63:554–569

    CAS  PubMed Central  PubMed  Google Scholar 

  • Palmer LK, Wolfe D, Keeley JL, Keil RL (2002) Volatile anesthetics affect nutrient availability in yeast. Genetics 161:563–574

    CAS  PubMed Central  PubMed  Google Scholar 

  • Palmer LK, Shoemaker JL, Wolfe D, Keil RL (2005) Inhibition of translation initiation by volatile anesthetics involves nutrient-sensitive GCN-independent and -dependent processes in yeast. Mol Biol Cell 16:3727–3739

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Palmer LK, Rannels SL, Kimball SR, Jefferson LS, Keil RL (2006) Inhibition of mammalian translation initiation by volatile anesthetics. Am J Physiol Endocrinol Metab 290:1267–1275

    Article  Google Scholar 

  • Pao SS, Paulsen IT, Saier MH Jr (1998) Major facilitator superfamily. Microbiol Mol Biol Rev 62:1–34

    CAS  PubMed Central  PubMed  Google Scholar 

  • Pringle MJ, Brown KB, Miller KW (1981) Can the lipid theories of anesthesia account for the cutoff in anesthetic potency in homologous series of alcohols? Mol Pharmacol 19:49–55

    CAS  PubMed  Google Scholar 

  • Prozialeck WC, Weiss BJ (1982) Inhibition of calmodulin by phenothiazines and related drugs: structure-activity relationships. Pharmacol Exp Ther 222:509–516

    CAS  Google Scholar 

  • Raghupathy E, Peterson NA, McKean CM (1970) Effects of phenothiazines on in vitro cerebral protein synthesis. Biochem Pharmacol 19:993–1000

    Article  CAS  Google Scholar 

  • Seeman P (1972) The membrane actions of anesthetics and tranquilizers. Pharmacol Rev 24:583–655

    CAS  PubMed  Google Scholar 

  • Sheth U, Parker R (2003) Decapping and decay of messenger RNA occur in cytoplasmic processing bodies. Science 300:805–808

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Smith ICP, Auger M, Jarrell HC (1991) Molecular details of anesthetic–lipid interaction. Ann NY Acad Sci 625:668–684

    Article  CAS  PubMed  Google Scholar 

  • Snyder SH, Banerjee SP, Yamamura HI, Greenburg D (1974) Drugs, neurotransmitters and schizophrenia. Science 184:1243–1253

    Article  CAS  PubMed  Google Scholar 

  • Sonenberg N, Hinnebusch AG (2009) Regulation of translation initiation in eukaryotes: mechanisms and biological targets. Cell 136:731–745

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Teixeira D, Parker R (2007) Analysis of P-body assembly in Saccharomyces cerevisiae. Mol Biol Cell 18:2274–2287

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Teixeira D, Sheth U, Valencia-Sanchez MA, Brengues M, Parker R (2005) Processing bodies require RNA for assembly and contain nontranslating mRNAs. RNA 11:371–382

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Toh-e A, Oguchi T (1998) Isolation and characterization of the yeast las21 mutants, which are sensitive to a local anestheticum, tetracaine. Genes Genet Syst 73:365–375

    Article  CAS  Google Scholar 

  • Toh-e A, Nakamura H, Oshima Y (1976) A gene controlling the synthesis of non-specific alkaline phosphatase in Saccharomyces cerevisiae. Biochim Biophys Acta 28:182–192

    Article  Google Scholar 

  • Uesono Y (2009) Environmental stresses and clinical drugs paralyze a cell. Commun Integr Biol 2:275–278

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Uesono Y, Toh-e A (2002) Transient inhibition of translation initiation by osmotic stress. J Biol Chem 277:13848–13855

    Article  CAS  PubMed  Google Scholar 

  • Uesono Y, Toh-e A, Kikuchi Y (1997) Ssd1p of Saccharomyces cerevisiae associates with RNA. J Biol Chem 272:16103–16109

    Article  CAS  PubMed  Google Scholar 

  • Uesono Y, Ashe MP, Toh-e A (2004) Simultaneous yet independent regulation of actin cytoskeletal organization and translation initiation by glucose in Saccharomyces cerevisiae. Mol Biol Cell 15:1544–1556

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Uesono Y, Araki T, Toh-e A (2008) Local anesthetics, antipsychotic phenothiazines, and cationic surfactants shut down intracellular reactions through membrane perturbation in yeast. Biosci Biotechnol Biochem 72:2884–2894

    Article  CAS  PubMed  Google Scholar 

  • Uesono Y, Toh-e A, Kikuchi Y, Terashima I (2011) Structural analysis of compounds with actions similar to local anesthetics and antipsychotic phenothiazines in yeast. Yeast 28:391–404

    Article  CAS  PubMed  Google Scholar 

  • Urban BW, Bleckwenn M, Barann M (2006) Interactions of anesthetics with their targets: non-specific, specific or both? Pharmacol Ther 111:729–770

    Article  CAS  PubMed  Google Scholar 

  • Wang D, Vo NV, Sowa GA, Hartman RA, Ngo K, Choe SR, Witt WT, Dong Q, Lee JY, Niedernhofer LJ, Kang JD (2011) Bupivacaine decreases cell viability and matrix protein synthesis in an intervertebral disc organ model system. Spine J 11:139–146

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wipf D, Ludewig U, Tegeder M, Rentsch D, Koch W, Frommer WB (2002) Conservation of amino acid transporters in fungi, plants and animals. Trends Biochem Sci 27:139–147

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Hiroyuki Hirano and Mitsutomo Abe for help with the protein detection system.

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Chihiro K. Watanabe, Takushi Hachiya, Ko Noguchi, Ichiro Terashima & Yukifumi Uesono

  2. Department of Molecular Biology, Saitama Medical School, 38 Morohongo, Moroyama, Iruma-gun, Saitama, 350-0495, Japan

    Tomoyuki Araki

  3. Medical Mycology Research Center (MMRC), Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8673, Japan

    Akio Toh-e

  4. Department of Life Science, Gakushuin University, Mejiro 1-5-1, Toshima-ku, Tokyo, 171-8588, Japan

    Yoshiko Kikuchi

Authors
  1. Tomoyuki Araki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Akio Toh-e
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoshiko Kikuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chihiro K. Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takushi Hachiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ko Noguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ichiro Terashima
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yukifumi Uesono
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yukifumi Uesono.

Additional information

Communicated by S. Hohmann.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Araki, T., Toh-e, A., Kikuchi, Y. et al. Tetracaine, a local anesthetic, preferentially induces translational inhibition with processing body formation rather than phosphorylation of eIF2α in yeast. Curr Genet 61, 43–53 (2015). https://doi.org/10.1007/s00294-014-0443-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00294-014-0443-0

Keywords

Search

Navigation