Identification and Characterization of the Three Members of the CLC Family of Anion Transport Proteins in Trypanosoma Brucei

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Dec 16

PMID 29244877

Citations 2

Authors

Michael E Steinmann

Remo S Schmidt

Juan P Macedo

Christina Kunz Renggli

Peter Butikofer

Doris Rentsch

Pascal Maser

Erwin Sigel

Affiliations

Soon will be listed here.

Abstract

CLC type anion transport proteins are homo-dimeric or hetero-dimeric with an integrated transport function in each subunit. We have identified and partially characterized three members of this family named TbVCL1, TbVCL2 and TbVCL3 in Trypanosoma brucei. Among the human CLC family members, the T. brucei proteins display highest similarity to CLC-6 and CLC-7. TbVCL1, but not TbVCL2 and TbVCL3 is able to complement growth of a CLC-deficient Saccharomyces cerevisiae mutant. All TbVCL-HA fusion proteins localize intracellulary in procyclic form trypanosomes. TbVCL1 localizes close to the Golgi apparatus and TbVCL2 and TbVCL3 to the endoplasmic reticulum. Upon expression in Xenopus oocytes, all three proteins induce similar outward rectifying chloride ion currents. Currents are sensitive to low concentrations of DIDS, insensitive to the pH in the range 5.4 to 8.4 and larger in nitrate than in chloride medium.

Citing Articles

Down the membrane hole: Ion channels in protozoan parasites.

Jimenez V, Mesones S PLoS Pathog. 2022; 18(12):e1011004.

PMID: 36580479 PMC: 9799330. DOI: 10.1371/journal.ppat.1011004.

Profilin is involved in G1 to S phase progression and mitotic spindle orientation during Leishmania donovani cell division cycle.

Ambaru B, Gangadharan G, Subramanya H, Gupta C PLoS One. 2022; 17(3):e0265692.

PMID: 35316283 PMC: 8939790. DOI: 10.1371/journal.pone.0265692.

References

Gonzalez-Salgado A, Steinmann M, Greganova E, Rauch M, Maser P, Sigel E . myo-Inositol uptake is essential for bulk inositol phospholipid but not glycosylphosphatidylinositol synthesis in Trypanosoma brucei. J Biol Chem. 2012; 287(16):13313-23. PMC: 3340000. DOI: 10.1074/jbc.M112.344812. View

GREENE J, Brown N, DiDomenico B, Kaplan J, Eide D . The GEF1 gene of Saccharomyces cerevisiae encodes an integral membrane protein; mutations in which have effects on respiration and iron-limited growth. Mol Gen Genet. 1993; 241(5-6):542-53. DOI: 10.1007/BF00279896. View

Braun N, Morgan B, Dick T, Schwappach B . The yeast CLC protein counteracts vesicular acidification during iron starvation. J Cell Sci. 2010; 123(Pt 13):2342-50. PMC: 2894126. DOI: 10.1242/jcs.068403. View

Jentsch T . Discovery of CLC transport proteins: cloning, structure, function and pathophysiology. J Physiol. 2015; 593(18):4091-109. PMC: 4594286. DOI: 10.1113/JP270043. View

Teste M, Duquenne M, Francois J, Parrou J . Validation of reference genes for quantitative expression analysis by real-time RT-PCR in Saccharomyces cerevisiae. BMC Mol Biol. 2009; 10:99. PMC: 2776018. DOI: 10.1186/1471-2199-10-99. View

Steinmann M, Gonzalez-Salgado A, Butikofer P, Maser P, Sigel E . A heteromeric potassium channel involved in the modulation of the plasma membrane potential is essential for the survival of African trypanosomes. FASEB J. 2015; 29(8):3228-37. DOI: 10.1096/fj.15-271353. View

Gaxiola R, Yuan D, Klausner R, Fink G . The yeast CLC chloride channel functions in cation homeostasis. Proc Natl Acad Sci U S A. 1998; 95(7):4046-50. PMC: 19960. DOI: 10.1073/pnas.95.7.4046. View

Estevez R, Pusch M, Ferrer-Costa C, Orozco M, Jentsch T . Functional and structural conservation of CBS domains from CLC chloride channels. J Physiol. 2004; 557(Pt 2):363-78. PMC: 1665104. DOI: 10.1113/jphysiol.2003.058453. View

Accardi A, Miller C . Secondary active transport mediated by a prokaryotic homologue of ClC Cl- channels. Nature. 2004; 427(6977):803-7. DOI: 10.1038/nature02314. View

10.

Wirtz E, Leal S, Ochatt C, Cross G . A tightly regulated inducible expression system for conditional gene knock-outs and dominant-negative genetics in Trypanosoma brucei. Mol Biochem Parasitol. 1999; 99(1):89-101. DOI: 10.1016/s0166-6851(99)00002-x. View

11.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

12.

Jentsch T, Steinmeyer K, Schwarz G . Primary structure of Torpedo marmorata chloride channel isolated by expression cloning in Xenopus oocytes. Nature. 1990; 348(6301):510-4. DOI: 10.1038/348510a0. View

13.

Ortiz D, Sanchez M, Quecke P, Landfear S . Two novel nucleobase/pentamidine transporters from Trypanosoma brucei. Mol Biochem Parasitol. 2008; 163(2):67-76. PMC: 2630410. DOI: 10.1016/j.molbiopara.2008.09.011. View

14.

Brenndorfer M, Boshart M . Selection of reference genes for mRNA quantification in Trypanosoma brucei. Mol Biochem Parasitol. 2010; 172(1):52-5. DOI: 10.1016/j.molbiopara.2010.03.007. View

15.

Huson D, Richter D, Rausch C, Dezulian T, Franz M, Rupp R . Dendroscope: An interactive viewer for large phylogenetic trees. BMC Bioinformatics. 2007; 8:460. PMC: 2216043. DOI: 10.1186/1471-2105-8-460. View

16.

Serricchio M, Butikofer P . Phosphatidylglycerophosphate synthase associates with a mitochondrial inner membrane complex and is essential for growth of Trypanosoma brucei. Mol Microbiol. 2012; 87(3):569-79. DOI: 10.1111/mmi.12116. View

17.

Sigel E, Minier F . The Xenopus oocyte: system for the study of functional expression and modulation of proteins. Mol Nutr Food Res. 2005; 49(3):228-34. DOI: 10.1002/mnfr.200400104. View

18.

Dohmen R, Strasser A, Honer C, Hollenberg C . An efficient transformation procedure enabling long-term storage of competent cells of various yeast genera. Yeast. 1991; 7(7):691-2. DOI: 10.1002/yea.320070704. View

19.

Picollo A, Liantonio A, Didonna M, Elia L, Conte Camerino D, Pusch M . Molecular determinants of differential pore blocking of kidney CLC-K chloride channels. EMBO Rep. 2004; 5(6):584-9. PMC: 1299079. DOI: 10.1038/sj.embor.7400169. View

20.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S . MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol. 2013; 30(12):2725-9. PMC: 3840312. DOI: 10.1093/molbev/mst197. View