Search results for "trna"
showing 3 items of 63 documents
Eukaryotic rRNA Modification by Yeast 5-Methylcytosine-Methyltransferases and Human Proliferation-Associated Antigen p120.
2015
International audience; Modified nucleotide 5-methylcytosine (m(5)C) is frequently present in various eukaryotic RNAs, including tRNAs, rRNAs and in other non-coding RNAs, as well as in mRNAs. RNA: m(5)C-methyltranferases (MTases) Nop2 from S. cerevisiae and human proliferation-associated nucleolar antigen p120 are both members of a protein family called Nop2/NSUN/NOL1. Protein p120 is well-known as a tumor marker which is over-expressed in various cancer tissues. Using a combination of RNA bisulfite sequencing and HPLC-MS/MS analysis, we demonstrated here that p120 displays an RNA:m(5)C-MTase activity, which restores m(5)C formation at position 2870 in domain V of 25S rRNA in a nop2 Delta …
Absolute quantification of noncoding RNA by microscale thermophoresis
2019
Abstract Accurate quantification of the copy numbers of noncoding RNA has recently emerged as an urgent problem, with impact on fields such as RNA modification research, tissue differentiation, and others. Herein, we present a hybridization‐based approach that uses microscale thermophoresis (MST) as a very fast and highly precise readout to quantify, for example, single tRNA species with a turnaround time of about one hour. We developed MST to quantify the effect of tRNA toxins and of heat stress and RNA modification on single tRNA species. A comparative analysis also revealed significant differences to RNA‐Seq‐based quantification approaches, strongly suggesting a bias due to tRNA modifica…
Caracterización bioquímica y funcional de MnmC, una proteína modificadora de tRNA de Escherichia coli
2015
En esta tesis doctoral se ha pretendido conocer aspectos bioquímicos y funcionales de la proteína bifuncional MnmC, modificadora de la U34 en algunos tRNAs del organismo Escherichia coli siempre teniendo en cuenta el marco de la ruta donde actúa (Ruta MnmEG). El primer objetivo fue clonar y expresar cada dominio de MnmC por separado para tener un sistema de estudio de las reacciones que ejercen por separado. Uno de los objetivos más importantes ha sido determinar cómo algunos de los tRNA sustratos de la ruta MnmEG son modificados por la enzima MnmC mediante un estudio comparativo de los dos dominios de la proteína y utilizando técnicas bioquímicas, genéticas y microbiólogicas para obtener d…