The RNA subunit of mitochondrial RNase P (mtP-RNA) is encoded by a mitochondrial gene (rnpB) in several ascomycete fungi and in the protists Reclinomonas americana and Nephroselmis olivacea. By searching for universally conserved structural elements, we have identified previously unknown rnpB genes in the mitochondrial DNAs (mtDNAs) of two fission yeasts, Schizosaccharomyces pombe and Schizosaccharomyces octosporus; in the budding yeast Pichia canadensis; and in the archiascomycete Taphrina deformans. The expression of mtP-RNAs of the predicted size was experimentally confirmed in the two fission yeasts, and their precise 5' and 3' ends were determined by sequencing of cDNAs generated from circularized mtP-RNAs. Comparative RNA secondary structure modeling shows that in contrast to mtP-RNAs of the two protists R. americana and N. olivacea, those of ascomycete fungi all have highly reduced secondary structures. In certain budding yeasts, such as Saccharomycopsis fibuligera, we find only the two most conserved pairings, P1 and P4. A P18 pairing is conserved in Saccharomyces cerevisiae and its close relatives, whereas nearly half of the minimum bacterial consensus structure is retained in the RNAs of fission yeasts, Aspergillus nidulans and Taphrina deformans. The evolutionary implications of the reduction of mtP-RNA structures in ascomycetes will be discussed.
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http://dx.doi.org/10.1261/rna.5880403 | DOI Listing |
BMC Res Notes
January 2025
Planta Piloto de Procesos Industriales Microbiológicos (PROIMI - CONICET), Tucumán, Argentina.
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View Article and Find Full Text PDFMethods Mol Biol
January 2025
Department of Immunobiology, University of Lausanne, Epalinges, Switzerland.
Fluorescence recovery after photobleaching (FRAP) can be employed to investigate membrane lipid mixing of vacuoles in live budding yeast cells and distinguish the fused, hemi-fused or non-fused states of these organelles under physiological conditions. Here, we describe a protocol for labeling the outer and inner leaflets of vacuoles in live cells that allow to detect hemifusion intermediates and, thus, identify components necessary for fusion pore opening.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, South Korea.
Cell-free in vitro assays offer several advantages for elucidating molecular mechanisms underlying various biological processes. Here, we describe a simple and quantitative in vitro assay using isolated yeast microsomes to measure homotypic ER membrane fusion. In this assay, membrane fusion between ER microsomes is monitored by reconstitution of luciferase activity from split luciferase fragments.
View Article and Find Full Text PDFMethods Mol Biol
January 2025
Dept of Biochemistry & Center for Biophysics and Quantitative Biology, University of Illinois Urbana-Champaign, Urbana, IL, USA.
Bio-Layer Interferometry (BLI) is a technique that uses optical biosensing to analyze interactions between molecules. The analysis of molecular interactions is measured in real-time and does not require fluorescent tags. BLI uses disposable biosensors that come in a variety of formats to bind different ligands including biotin, hexahistidine, GST, and the Fc portion of antibodies.
View Article and Find Full Text PDFSci Rep
January 2025
Rui Pu Agricultural Technology Co., Ltd, Hohhot, Inner Mongolia, People's Republic of China.
Duddingtonia flagrans is a nematode-trapping fungus that is widely used to control parasitic nematodes in livestock. After oral ingestion and passage through the digestive tract of animals, this microorganism captures nematodes in feces. Although many researchers have examined the safety of this fungus for humans, animals, and the environment, few reports have discussed the safety of nematode-trapping D.
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