Pentatricopeptide repeat proteins in Trypanosoma brucei function in mitochondrial ribosomes.
ABSTRACT The pentatricopeptide repeat (PPR), a degenerate 35-amino-acid motif, defines a novel eukaryotic protein family. Plants have 400 to 500 distinct PPR proteins, whereas other eukaryotes generally have fewer than 5. The few PPR proteins that have been studied have roles in organellar gene expression, probably via direct interaction with RNA. Here we show that the parasitic protozoan Trypanosoma brucei encodes 28 distinct PPR proteins, an extraordinarily high number for a nonplant organism. A comparative analysis shows that seven out of eight selected PPR proteins are mitochondrially localized and essential for oxidative phosphorylation. Six of these are required for the stabilization of mitochondrial rRNAs and, like ribosomes, are associated with the mitochondrial membranes. Furthermore, one of the PPR proteins copurifies with the large subunit rRNA. Finally, ablation of all of the PPR proteins that were tested induces degradation of the other PPR proteins, indicating that they function in concert. Our results show that a significant number of trypanosomal PPR proteins are individually essential for the maintenance and/or biogenesis of mitochondrial rRNAs.
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ABSTRACT: The conjugacy of saccades is rapidly modified if the images are made unequal for the two eyes. Disconjugacy persists even in the absence of disparity which indicates learning. Binocular visual disparity is a major cue to depth and is believed to drive the disconjugacy of saccades to aniseikonic images. The goal of the present study was to test whether monocular depth cues can also influence the disconjugacy of saccades. Three experiments were performed in which subjects were exposed for 15-20 min to a 10% image size inequality. Three different images were used: a grid that contained a single monocular depth cue strongly indicating a frontoparallel plane; a random-dot pattern that contained a less prominent monocular depth cue (absence of texture gradient) which also indicates the frontoparallel plane; and a complex image with several overlapping geometric forms that contained a variety of monocular depth cues. Saccades became disconjugate in all three experiments. The disconjugacy was larger and more persistent for the experiment using the random-dot pattern that had the least prominent monocular depth cues. The complex image which had a large variety of monocular depth cues produced the most variable and less persistent disconjugacy. We conclude that the monocular depth cues modulate the disconjugacy of saccades stimulated by the disparity of aniseikonic images.Vision Research 10/1999; 39(18):3109-22. · 2.41 Impact Factor
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ABSTRACT: African trypanosomes, such as Trypanosoma brucei, are protozoan parasites of mammals that were first described over 100 hundred years ago. They have long been the subjects of biological investigation, which has yielded insights into a number of fundamental, as well as novel, cellular processes in all organisms. In the last decade or so, genetic manipulation of trypanosomes has become possible through DNA transformation, allowing yet more detailed analysis of the biology of the parasite. One facet of this is that DNA transformation has itself been used as an assay for recombination and will undoubtedly lead to further genetic approaches to examine this process. Here we describe protocols for DNA transformation of Trypanosoma brucei, including two different life cycle stages and two different strain types that are distinguished by morphological and developmental criteria. We consider the application of transformation to recombination, as well as the uses of transforming the different life cycle stages and strain types.Methods in molecular biology (Clifton, N.J.) 02/2004; 262:53-86.
Article: Isolation and characterization of mitochondrial ribosomes and ribosomal subunits from Leishmania tarentolae.[show abstract] [hide abstract]
ABSTRACT: We have analyzed Leishmania tarentolae mitochondrial ribonucleoprotein (RNP) complexes using the 9S small subunit (SSU) rRNA and the 12S large subunit (LSU) rRNA as markers, and have identified a 50S RNP particle as the putative mitochondrial monosome, a 40S particle as the putative LSU and a 30S particle as the putative SSU. These assignments are supported by morphological analysis by cryo-electron microscopy and proteomics analyses by mass spectrometry. The presence of additional rRNA-containing particles complicated the analysis and most likely was the basis for previous difficulties in identification of these ribosomes; thus, in addition to the monosomes and their subunits, there are abundant stable 45S particles (SSU(*)) containing only 9S rRNA, which may represent homodimers of the SSU or SSU associated with additional proteins, and variable minor amounts of 65S and 70S particles, which represent homodimers of the LSU and SSU(*), respectively. These additional rRNA particles might be due to the lengthy mitochondrial isolation and ribosome isolation procedures or may be present in vivo and play yet undetermined roles.Molecular and Biochemical Parasitology 08/2006; 148(1):69-78. · 2.55 Impact Factor