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Molecular and Cellular Biology, November 2008, p. 6939-6953, Vol. 28, No. 22
0270-7306/08/$08.00+0     doi:10.1128/MCB.01115-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

The KREPA3 Zinc Finger Motifs and OB-Fold Domain Are Essential for RNA Editing and Survival of Trypanosoma brucei{triangledown}

Xuemin Guo,1 Nancy Lewis Ernst,1 and Kenneth D. Stuart1,2*

Seattle Biomedical Research Institute, Seattle, Washington 98109,1 Department of Pathobiology, University of Washington, Seattle, Washington 981952

Received 15 July 2008/ Returned for modification 14 August 2008/ Accepted 3 September 2008

Three types of editosomes, each with an identical core containing six related KREPA proteins, catalyze the U insertion and deletion RNA editing of mitochondrial mRNAs in trypanosomes. Repression of expression of one of these, KREPA3 (also known as TbMP42), shows that it is essential for growth and in vivo editing in both procyclic (PF) and bloodstream (BF) life cycle stages of Trypanosoma brucei. RNA interference knockdown results in editosome disruption and altered in vitro editing in PFs, while repression by regulatable double knockout results in almost complete loss of editosomes in BFs. Mutational analysis shows that the KREPA3 zinc fingers and OB-fold domain are each essential for growth and in vivo editing. Nevertheless, KREPA3 with mutated zinc fingers incorporates into editosomes that catalyze in vitro editing and thus is not essential for editosome integrity, although stability is affected. In contrast, the OB-fold domain is essential for editosome integrity. Overall, KREPA3, especially its OB-fold, functions in editosome integrity, and its zinc fingers are essential for editing in vivo but not for the central catalytic steps. KREPA3 may function in editosome organization and/or RNA positioning.

* Corresponding author. Mailing address: Seattle Biomedical Research Institute, 307 Westlake Avenue North, Suite 500, Seattle, WA 98109. Phone: (206) 256-7316. Fax: (206) 256-7229. E-mail: ken.stuart{at}sbri.org

{triangledown} Published ahead of print on 15 September 2008.

Molecular and Cellular Biology, November 2008, p. 6939-6953, Vol. 28, No. 22
0270-7306/08/$08.00+0     doi:10.1128/MCB.01115-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ernst, N. L., Panicucci, B., Carnes, J., Stuart, K. (2009). Differential functions of two editosome exoUases in Trypanosoma brucei. RNA 15: 947-957 [Abstract] [Full Text]  
  • Niemann, M., Kaibel, H., Schluter, E., Weitzel, K., Brecht, M., Goringer, H. U. (2009). Kinetoplastid RNA editing involves a 3' nucleotidyl phosphatase activity. Nucleic Acids Res 37: 1897-1906 [Abstract] [Full Text]  


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