Abstract
The presumptive choline transporter, CTL1, was initially identified through functional complementation of a triple yeast mutant (ctr ise URA3Δ) with deficiencies in both choline transport and choline neosynthesis under selective conditions that cause perturbations in membrane synthesis and growth. After transformation of these yeasts with a heterologous yeast expression library made from Torpedo electric lobe cDNAs, several colonies showed increased growth but only one clone increased the accumulation of external choline. The corresponding full-length cDNA was isolated and encodes a protein with 10 transmembrane domains. Northern analysis of Torpedo mRNA indicates that CTL1 is expressed at high levels in the spinal cord and brain. In Xenopus oocytes, Torpedo CTL1 expression was associated with the appearance of sodium independent high-affinity choline uptake. We propose that CTL1 plays a role in providing choline for membrane synthesis in the nervous system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
REFERENCES
Nikawa, J., Hosaka, K., Tsukagoshi, Y., and Yamashita, S. 1990. Primary structure of the yeast choline transport gene and regulation of its expression. J. Biol. Chem. 265:15996–16003.
Andresen, P. A., Kaasen, I., Styrvold, O. B., Boulnois, G., and Strom, A. R. 1988. Molecular cloning, physical mapping and expression of the bet genes governing the osmoregulatory choline-glycine betaine pathway of Escherichia coli. J. Gen. Microbiol. 134:1737–1746.
Sweet, D. H., Miller, D. S., and Pritchard, J. B. 2001. Ventricular choline transport: A role for organic cation transporter 2 expressed in choroid plexus. J. Biol. Chem. 276:41611–41619.
Simon, J. R. and Kuhar, M. J. 1976. High affinity choline uptake: Ionic and energy requirements. J. Neurochem. 27:93–99.
Okuda, T., Haga, T., Kanai, Y., Endou, H., Ishihara, T., and Katsura, I. 2000. Identification and characterization of the high-affinity choline transporter. Nat. Neurosci. 3:120–125.
Misawa, H., Nakata, K., Matsuura, J., Nagao, M., Okuda, T., and Haga, T. 2001. Distribution of the high-affinity choline transporter in the central nervous system of the rat. Neuroscience 105:87–98.
Tuček, S. 1984. Problems in the organization and control of acetylcholine synthesis in brain neurons. Prog. Biophys. Mol. Biol 44:1–46.
Martin, K. 1968. Concentrative accumulation of choline by human erythrocytes. J. Gen. Physiol. 51:497–516.
Massarelli, R., Ciesielski-Treska, J., Ebel, A., and Mandel, P. 1974. Choline uptake in glial cell cultures. Brain Res. 81:361–363.
O'Regan, S., Traiffort, E., Ruat, M., Cha, N., Compaore, D., and Meunier, F. M. 2000. An electric lobe suppressor for a yeast choline transport mutation belongs to a new family of transporter-like proteins. Proc. Natl. Acad. Sci. USA 97:1835–1840.
Campbell, I. 1988. Standard media for cultivation of yeasts. Pages 277–280, in Campbell, I. and Duffus, J. H. (ed.), Yeast: A Practical Approach, IRL Press, Oxford.
Minet, M., Dufour, M. E., and Lacroute, F. 1992. Complementation of Saccharomyces cerevisiae auxotrophic mutants by Arabidopsis thaliana cDNAs. Plant J. 2:417–422.
Gietz, R. D., Schiestl, R. H., Willems, A. R., and Woods, R. A. 1995. Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360.
O'Regan, S., Birman, S., and Meunier, F. M. 1995. Regulation of hemicholinium-3 sensitive choline uptake in Xenopus laevis oocytes by the second C2 domain of synaptotagmin. Brain Res. Mol. Brain Res. 32:135–142.
Choy, P. C., Whitehead, F. W., and Vance, D. E. 1978. A rapid method for the determination of CTP and phosphocholine in rat liver and baby hamster kidney 21 cells. Can. J. Biochem. 56:831–835.
Huff, T., Muller, C. S., Otto, A. M., Netzker, R., and Hannappel, E. 2001. Beta-thymosins, small acidic peptides with multiple functions. Int. J. Biochem. Cell Biol. 33:205–220.
Serru, V., Le Naour, F., Billard, M., Azorsa, D. O., Lanza, F., Boucheix, C., and Rubinstein, E. 1999. Selective tetraspan-integrin complexes (CD81/alpha4beta1, CD151/alpha3beta1, CD151/alpha6beta1) under conditions disrupting tetraspan interactions. Biochem. J. 340:103–111.
Ledesma, M. D., Simons, K., and Dotti, C. G. 1998. Neuronal polarity: Essential role of protein-lipid complexes in axonal sorting. Proc. Natl. Acad. Sci. USA 95:3966–3971.
Wille, S., Szekeres, A., Majdic, O., Prager, E., Staffler, G., Stockl, J., Kunthalert, D., Prieschl, E. E., Baumruker, T., Burtscher, H., Zlabinger, G. J., Knapp, W., and Stockinger, H. 2001. Characterization of CDw92 as a member of the choline transporter-like protein family regulated specifically on dendritic cells. J. Immunol. 167:5795–5804.
Blusztajn, J. K. 1998. Choline: A vital amine. Science 281:794–795.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
O'Regan, S., Meunier, FM. Selection and Characterization of the Choline Transport Mutation Suppressor from Torpedo Electric Lobe, CTL1. Neurochem Res 28, 551–555 (2003). https://doi.org/10.1023/A:1022877524469
Issue Date:
DOI: https://doi.org/10.1023/A:1022877524469