Abstract
This chapter describes the methods used to purify the RNA-editing complex, to identify the proteins by mass spectrometry, and to demonstrate the functions for some of the proteins.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kable, M. L., Heidmann, S., and Stuart, K. (1997) RNA editing: getting U into RNA. Trends Biochem. Sci. 22, 162–166.
Stuart, K. and Panigrahi, A. K. (2002) RNA editing: complexity and complications. Mol. Microbiol. 45, 591–596.
Simpson, L., Sbicego, S., and Aphasizhev, R. (2003) Uridine insertion/deletion RNA editing in trypanosome mitochondria: a complex business. RNA 9, 265–276.
Panigrahi, A. K., Gygi, S., Ernst, N., (2001) Association of two novel proteins, TbMP52 and TbMP48, with the Trypanosoma brucei RNA editing complex. Mol. Cell. Biol. 21, 380–389.
Panigrahi, A. K., Schnaufer, A., Carmean, N., et al. (2001) Four related proteins of the T. brucei RNA editing complex. Mol. Cell. Biol. 21, 6833–6840.
Panigrahi, A. K., Schnaufer, A., Ernst, N., Wang, B., Carmean, N., Salavati, R., and Stuart, K. (2003) Identification of novel components of Trypanosoma brucei editosome. RNA 9, 484–492.
Stuart, K., Gobright, E., Jenni, L., Milhausen, M., Thomashow, L., and Agabian, N. (1984) The IsTaR 1 serodeme of Trypanosoma brucei: development of a new serodeme. J. Parasitol. 70, 747–754.
Harris, M. E., Moore, D. R., and Hajduk, S. L. (1990) Addition of uridines to edited RNAs in trypanosome mitochondria occurs independently of transcription. J. Biol. Chem. 265, 11,368–11,376.
Rigaut, G., Shevchenko, A., Rutz, B., Wilm, M., Mann, M., and Seraphin, B. (1999) A generic protein purification method for protein complex characterization and proteome exploration. Nat. Biotechnol. 17, 1030–1032.
Estévez, A. M., Kempf, T., and Clayton, C. (2001) The exosome of Trypanosoma brucei. EMBO J. 20, 3831–3839.
Wirtz, E., Simone, L., Claudia, O., and Cross, G. A. M. (1999) A tightly regulated inducible expression system for conditional gene knock-outs and dominant-negative genetics in Trypanosoma brucei. Mol. Biochem. Parasitol. 99, 89–101.
Sambrook, J., Fritsch, T., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Brun, R. and Schonenberger, M. (1979) Cultivation and in vitro cloning of procyclic culture forms of Trypanosoma brucei in a semi-defined medium. Acta Trop. 36, 289–292.
Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68, 850–858.
Panigrahi, A. K., Allen, T. E., Haynes, P. A., Gygi, S. P., and Stuart, K. (2003) Mass spectrometric analysis of the editosome and other multiprotein complexes in Trypanosoma brucei. J. Am. Soc. Mass Spectrom. 14, 728–735.
Schnaufer, A., Panigrahi, A. K., Panicucci, B., Igo, R. P. Jr., Salavati, R., and Stuart, K. D. (2001) An RNA ligase essential for RNA editing and survival of the bloodstream form of Trypanosoma brucei. Science 291, 2159–2162.
Palazzo, S. S., Panigrahi, A. K., Igo, R. P. Jr., Salavati, R., and Stuart, K. (2003) Kinetoplastid RNA editing ligases: complex association, characterization, and substrate requirements. Mol. Biochem. Parasitol. 127, 161–167.
Ernst, N., Igo, R. P. Jr., Panicucci, B., Panigrahi, A. K., and Stuart, K. (2003) TbMP57 is a 3′ terminal uridylyl transferase (TUTase) of the Trypanosoma brucei editosome. Mol. Cell 11, 1525–1536.
Drozdz, M., Palazzo, S. S., Salavati, R., O’Rear, J., Clayton, C., and Stuart, K. (2002) TbMP81 is required for RNA editing in iTrypanosoma brucei. EMBO J. 21, 1791–1799.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc.
About this protocol
Cite this protocol
Stuart, K., Panigrahi, A.K., Schnaufer, A. (2004). Identification and Characterization of Trypanosome RNA-Editing Complex Components. In: Gott, J.M. (eds) RNA Interference, Editing, and Modification. Methods in Molecular Biology, vol 265. Humana Press. https://doi.org/10.1385/1-59259-775-0:273
Download citation
DOI: https://doi.org/10.1385/1-59259-775-0:273
Publisher Name: Humana Press
Print ISBN: 978-1-58829-242-1
Online ISBN: 978-1-59259-775-8
eBook Packages: Springer Protocols