Trypanosoma brucei mitochondrial guide RNA-mRNA chimera-forming activity cofractionates with an editing-domain-specific endonuclease and RNA ligase and is mimicked by heterologous nuclease and RNA ligase.

Molecular and Cellular Biology
K J PillerB Sollner-Webb

Abstract

RNA editing in trypanosomes has been proposed to occur through transesterification or endonuclease cleavage and RNA ligation reactions. Both models involve a chimeric intermediate in which a guide RNA (gRNA) is joined through its 3' oligo(U) tail to an editing site of the corresponding mRNA. Velocity centrifugation of Trypanosoma brucei mitochondrial extracts had been reported to completely separate the gRNA-mRNA chimera-forming activity from endonuclease activity (V. W. Pollard, M. E. Harris, and S. L. Hajduk, EMBO J. 11:4429-4438, 1992), appearing to rule out the endonuclease-RNA ligase mechanism. However, we show that an editing-domain-specific endonuclease activity does cosediment with the chimera-forming activity, as does the RNA ligase activity, but detection of the specific endonuclease requires reducing assay conditions. This report further demonstrates that the T. brucei chimera-forming activity is mimicked by mung bean nuclease and T4 RNA ligase. Using cytochrome b (CYb) preedited mRNA and a model CYb gRNA, we found that these heterologous enzymes specifically generate CYb gRNA-mRNA chimeras analogous to those formed in the mitochondrial extract. These combined results provide support for the endonuclease-RNA ligase m...Continue Reading

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Citations

Jul 31, 2007·Methods in Enzymology·Jason Carnes, Kenneth D Stuart
Nov 5, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jason CarnesKenneth Stuart
Feb 7, 1997·The Journal of Biological Chemistry·J KöllerH U Göringer
Feb 13, 2002·Molecular and Cellular Biology·Robert P IgoKenneth Stuart
Apr 1, 1996·Current Opinion in Genetics & Development·R Benne
Jun 3, 1996·Biochimica Et Biophysica Acta·G J Arts, R Benne
Jul 1, 1997·Molecular and Cellular Biology·F BringaudL Simpson
Jun 1, 1995·Molecular and Cellular Biology·K J PillerB Sollner-Webb
Mar 1, 1997·Microbiology and Molecular Biology Reviews : MMBR·K StuartS D Seiwert
Aug 30, 1996·Science·B Sollner-Webb

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African trypanosomiasis, also known as sleeping sickness, is an insect-borne parasitic disease of humans and other animals. It is caused by protozoa of the species Trypanosoma brucei and almost invariably progresses to death unless treated. Discover the latest research on African trypanosomiasis here.