Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (604K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (29)
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Blom, D.
Right arrow Articles by Benne, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Blom, D.
Right arrow Articles by Benne, R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 14 2950-2962
© 2001 Oxford University Press

Cloning and characterization of two guide RNA-binding proteins from mitochondria of Crithidia fasciculata: gBP27, a novel protein, and gBP29, the orthologue of Trypanosoma brucei gBP21

Daniël Blom, Janny van den Burg, Cornelis K. D. Breek, Dave Speijer, Anton O. Muijsers and Rob Benne*

Department of Biochemistry, Academic Medical Centre, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands

In kinetoplastid protozoa, mitochondrial (mt) mRNAs are post-transcriptionally edited by insertion and deletion of uridylate residues, the information being provided by guide (g)RNAs. Currently popular mechanisms for the editing process envisage a series of consecutive ‘cut-and-paste’ reactions, carried out by a complex RNP machinery. Here we report on the purification, cloning and functional analysis of two gRNA-binding proteins of 28.8 (gBP29) and 26.8 kDa (gBP27) from mitochondria of the insect trypanosome Crithidia fasciculata. gBP29 and gBP27 proved to be similar, Arg + Ala-rich proteins, with pI values of ~10.0. gBP27 has no homology to known proteins, but gBP29 is the C.fasciculata orthologue of gBP21 from Trypanosoma brucei, a gRNA-binding protein that associates with active RNA editing complexes. As measured in UV cross-linking assays, His-tagged recombinant gBP29 and gBP27 bind to radiolabelled poly(U) and synthetic gRNAs, while competition experiments suggest a role for the gRNA 3'-(U)-tail in binding to these proteins. Immunoprecipitates of mt extracts generated with antibodies against gBP29 also contained gBP27 and vice versa. The immunoprecipitates further harbored a large proportion of the cellular content of four different gRNAs and of edited and pre-edited NADH dehydrogenase subunit 7 mRNAs, but only small amounts of mt rRNAs. In addition, the bulk of gBP29 and gBP27 co-eluted with gRNAs from gel filtration columns in the high molecular weight range. Together, these results suggest that the proteins are part of a large macromolecular complex(es). We infer that gBP29 and gBP27 are components of the C.fasciculata editing machinery that may interact with gRNAs.

* To whom correspondence should be addressed. Tel: +1 31 20 5665131; Fax: +1 31 20 6915519; Email: r.benne{at}amc.uva.nlPresent address:Cornelis K. D. Breek, Institute of Plant Molecular Sciences, Clusius Laboratory, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands +AF156855, AF157559, AF373808


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
M. Niemann, M. Brecht, E. Schluter, K. Weitzel, M. Zacharias, and H. U. Goringer
TbMP42 is a structure-sensitive ribonuclease that likely follows a metal ion catalysis mechanism
Nucleic Acids Res., August 1, 2008; 36(13): 4465 - 4473.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
M. L. Ammerman, J. C. Fisk, and L. K. Read
gRNA/pre-mRNA annealing and RNA chaperone activities of RBP16
RNA, June 1, 2008; 14(6): 1069 - 1080.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Cifuentes-Rojas, P. Pavia, A. Hernandez, D. Osterwisch, C. Puerta, and J. Cruz-Reyes
Substrate Determinants for RNA Editing and Editing Complex Interactions at a Site for Full-round U Insertion
J. Biol. Chem., February 16, 2007; 282(7): 4265 - 4276.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
A. Sacharidou, C. Cifuentes-Rojas, K. Halbig, A. Hernandez, L. J. Dangott, M. De Nova-Ocampo, and J. Cruz-Reyes
RNA editing complex interactions with a site for full-round U deletion in Trypanosoma brucei
RNA, July 1, 2006; 12(7): 1219 - 1228.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
C. Cifuentes-Rojas, K. Halbig, A. Sacharidou, M. De Nova-Ocampo, and J. Cruz-Reyes
Minimal pre-mRNA substrates with natural and converted sites for full-round U insertion and U deletion RNA editing in trypanosomes
Nucleic Acids Res., November 23, 2005; 33(20): 6610 - 6620.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
G. Gao, A. M. Simpson, X. Kang, K. Rogers, M. Nebohacova, F. Li, and L. Simpson
Functional complementation of Trypanosoma brucei RNA in vitro editing with recombinant RNA ligase
PNAS, March 29, 2005; 102(13): 4712 - 4717.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
E. Vondruskova, J. van den Burg, A. Zikova, N. L. Ernst, K. Stuart, R. Benne, and J. Lukes
RNA Interference Analyses Suggest a Transcript-specific Regulatory Role for Mitochondrial RNA-binding Proteins MRP1 and MRP2 in RNA Editing and Other RNA Processing in Trypanosoma brucei
J. Biol. Chem., January 28, 2005; 280(4): 2429 - 2438.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
J. L. Penschow, D. A. Sleve, C. M. Ryan, and L. K. Read
TbDSS-1, an Essential Trypanosoma brucei Exoribonuclease Homolog That Has Pleiotropic Effects on Mitochondrial RNA Metabolism
Eukaryot. Cell, October 1, 2004; 3(5): 1206 - 1216.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
L. SIMPSON, R. APHASIZHEV, G. GAO, and X. KANG
Mitochondrial proteins and complexes in Leishmania and Trypanosoma involved in U-insertion/deletion RNA editing
RNA, February 1, 2004; 10(2): 159 - 170.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
B. Wang, N. L. Ernst, S. S. Palazzo, A. K. Panigrahi, R. Salavati, and K. Stuart
TbMP44 Is Essential for RNA Editing and Structural Integrity of the Editosome in Trypanosoma brucei
Eukaryot. Cell, June 1, 2003; 2(3): 578 - 587.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
M. PELLETIER and L. K. READ
RBP16 is a multifunctional gene regulatory protein involved in editing and stabilization of specific mitochondrial mRNAs in Trypanosoma brucei
RNA, April 1, 2003; 9(4): 457 - 468.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
A. K. PANIGRAHI, A. SCHNAUFER, N. L. ERNST, B. WANG, N. CARMEAN, R. SALAVATI, and K. STUART
Identification of novel components of Trypanosoma brucei editosomes
RNA, April 1, 2003; 9(4): 484 - 492.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
L. SIMPSON, S. SBICEGO, and R. APHASIZHEV
Uridine insertion/deletion RNA editing in trypanosome mitochondria: A complex business
RNA, March 1, 2003; 9(3): 265 - 276.
[Abstract] [Full Text] [PDF]

Home page
 RNAHome page
R. APHASIZHEV, I. APHASIZHEVA, R. E. NELSON, and L. SIMPSON
A 100-kD complex of two RNA-binding proteins from mitochondria of Leishmania tarentolae catalyzes RNA annealing and interacts with several RNA editing components
RNA, January 1, 2003; 9(1): 62 - 76.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
C. E. Huang, S. F. O'Hearn, and B. Sollner-Webb
Assembly and Function of the RNA Editing Complex in Trypanosoma brucei Requires Band III Protein
Mol. Cell. Biol., May 1, 2002; 22(9): 3194 - 3203.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
U. F. Muller and H. U. Goringer
Mechanism of the gBP21-mediated RNA/RNA annealing reaction: matchmaking and charge reduction
Nucleic Acids Res., January 15, 2002; 30(2): 447 - 455.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.