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J. Biol. Chem., Vol. 277, Issue 49, 47348-47357, December 6, 2002
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From the Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan
52900, Israel and the Trypanosomes are protozoan parasites that have a
major impact on health. This family diverged very early from the
eukaryotic lineage and possesses unique RNA processing mechanisms such
as trans-splicing and RNA editing. The trypanosome signal
recognition particle (SRP) has a unique composition compared with all
known SRP complexes, because it contains two RNA molecules, the 7SL RNA
and a tRNA-like molecule. RNA interference was utilized to elucidate
the essentiality of the SRP pathway and its role in protein
translocation in Trypanosoma brucei. The production of double stranded RNA specific for the signal peptide-binding
protein SRP54 induced the degradation of the mRNA and a loss of the
SRP54 protein. SRP54 depletion elicited inhibition in growth and
cytokinesis, suggesting that the SRP pathway is essential. The
translocation of four signal peptide-containing proteins was examined.
Surprisingly, the proteins were translocated to the endoplasmic
reticulum and properly processed. However, the surface EP procyclin,
the lysosomal protein p67, and the flagellar pocket protein CRAM were
mislocalized and accumulated in megavesicles, most likely because of a
secondary effect on protein sorting. The translocation of these
proteins to the endoplasmic reticulum under SRP54 depletion suggests
that an alternative pathway for protein translocation exists in trypanosomes.
RNA Interference of Signal Peptide-binding Protein SRP54 Elicits
Deleterious Effects and Protein Sorting Defects in Trypanosomes*
, and Department of Medicine and Cell
Biology, Yale University School of Medicine,
New Haven, Connecticut 06520-8022
*
This work was supported in part by a grant from the Israel
Science Foundation.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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