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Molecular and Cellular Biology, January 2001, p. 380-389, Vol. 21, No. 2
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.2.380-389.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Association of Two Novel Proteins, TbMP52 and
TbMP48, with the Trypanosoma brucei RNA Editing
Complex
Aswini K.
Panigrahi,1,2
Steven P.
Gygi,3
Nancy L.
Ernst,1,2
Robert P.
Igo Jr.,1,2
Setareh S.
Palazzo,1,2
Achim
Schnaufer,1,2
David S.
Weston,1,2
Nicole
Carmean,1
Reza
Salavati,1,2
Ruedi
Aebersold,3 and
Kenneth D.
Stuart1,2,*
Seattle Biomedical Research Institute,
Seattle, Washington 98109,1 and Departments of
Pathobiology2 and Molecular
Biotechnology,3 University of Washington,
Seattle, Washington 98195
Received 3 August 2000/Returned for modification 29 September
2000/Accepted 19 October 2000
RNA editing in kinetoplastid mitochondria inserts and deletes
uridylates at multiple sites in pre-mRNAs as directed by guide RNAs.
This occurs by a series of steps that are catalyzed by
endoribonuclease, 3'-terminal uridylyl transferase, 3'-exouridylylase,
and RNA ligase activities. A multiprotein complex that contains these
activities and catalyzes deletion editing in vitro was enriched from
Trypanosoma brucei mitochondria by sequential
ion-exchange and gel filtration chromatography, followed by glycerol
gradient sedimentation. The complex size is approximately 1,600 kDa,
and the purified fraction contains 20 major polypeptides. A monoclonal
antibody that was generated against the enriched complex reacts with an
~49-kDa protein and specifically immunoprecipitates in vitro
deletion RNA editing activity. The protein recognized by the antibody
was identified by mass spectrometry, and the corresponding gene,
designated TbMP52, was cloned. Recombinant TbMP52
reacts with the monoclonal antibody. Another novel protein, TbMP48,
which is similar to TbMP52, and its gene were also identified in the
enriched complex. These results suggest that TbMP52 and TbMP48 are
components of the RNA editing complex.
*
Corresponding author. Mailing address: Seattle
Biomedical Research Institute, 4 Nickerson St., Seattle, WA 98109. Phone: (206) 284-8846, ext. 316. Fax: (206) 284-0313. E-mail:
kstuart{at}u.washington.edu.
Molecular and Cellular Biology, January 2001, p. 380-389, Vol. 21, No. 2
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.2.380-389.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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