ABSTRACT: mTRAN proteins are are plant specific-components of the mitoribosome. To asses of loss-of-function mtran1 mtran2 double mutants show decreased translation rates, we performed ribosome footprinting coupled to RNA-seq (Ribo-seq).
Project description:mTRAN proteins are components of the plant mitochondrial small subunits, thought to bind the mRNA 5' regions to initiate translation. This experiment was designed to identify the mTRAN1 binding regions on the plant mitochondrial mRNAs
Project description:Aiming at defining the protein content and composition of a single mitochondrion of Arabidopsis thaliana, shotgun analyses were performed on purified mitochondria from cultured heterotrophic Arabidopsis cells. Based on microscopic observations on the size of mitochondria, as well as biochemical properties, such as protein concentration, average molecular mass of proteins, and the average length of the signal peptide, intensity-based absolute quantification (iBAQ) values were applied to calculate the copy number of each identified protein species within a single mitochondrion. Copy numbers of the individual proteins span five orders of magnitude, ranging from >40,000 for Voltage-Dependent Anion Channel 1 (VDAC1) to sub-stoichiometric copy numbers, i.e. less than a single copy per single mitochondrion, for several pentatricopeptide repeat (PPR) proteins that modify mitochondrial transcripts. Aiming at defining the protein content and composition of a single mitochondrion of Arabidopsis thaliana, shotgun analyses were performed on purified mitochondria from cultured heterotrophic Arabidopsis cells. Based on microscopic observations on the size of mitochondria, as well as biochemical properties, such as protein concentration, average molecular mass of proteins, and the average length of the signal peptide, intensity-based absolute quantification (iBAQ) values were applied to calculate the copy number of each identified protein species within a single mitochondrion. Copy numbers of the individual proteins span five orders of magnitude, ranging from >40,000 for Voltage-Dependent Anion Channel 1 (VDAC1) to sub-stoichiometric copy numbers, i.e. less than a single copy per single mitochondrion, for several pentatricopeptide repeat (PPR) proteins that modify mitochondrial transcripts. Aiming at defining the protein content and composition of a single mitochondrion of Arabidopsis thaliana, shotgun analyses were performed on purified mitochondria from cultured heterotrophic Arabidopsis cells. Based on microscopic observations on the size of mitochondria, as well as biochemical properties, such as protein concentration, average molecular mass of proteins, and the average length of the signal peptide, intensity-based absolute quantification (iBAQ) values were applied to calculate the copy number of each identified protein species within a single mitochondrion. Copy numbers of the individual proteins span five orders of magnitude, ranging from >40,000 for Voltage-Dependent Anion Channel 1 (VDAC1) to sub-stoichiometric copy numbers, i.e. less than a single copy per single mitochondrion, for several pentatricopeptide repeat (PPR) proteins that modify mitochondrial transcripts.
Project description:Genomic DNA from 305 Col x Ct F2 individuals was extracted by CTAB and used to generate sequencing libraries as previously described (Hennig et al, 2018 G3). Sequencing data was analysed to identify crossovers using the TIGER pipeline as previously described (Rowan et al, 2015 G3; Yelina et al, 2015 Genes & Dev).
Project description:Genomic DNA from 320 msh2 Col x Ct F2 individuals was extracted by CTAB and used to generate sequencing libraries as previously described (Hennig et al, 2018 G3). Sequencing data was analysed to identify crossovers using the TIGER pipeline as previously described (Rowan et al, 2015 G3; Yelina et al, 2015 Genes & Dev).
Project description:Genomic DNA from 241 sni1-/- Col x Ct F2 individuals was extracted by CTAB and used to generate sequencing libraries as previously described (Hennig et al, 2018 G3). Sequencing data was analysed to identify crossovers using the TIGER pipeline as previously described (Rowan et al, 2015 G3; Yelina et al, 2015 Genes & Dev).
Project description:Light is the most important cue for plant metabolism since its presence enables photosynthesis. Mitochondria respond to light conditions by adjusting flow through the citric acid cycle and the respiratory chain to support photosynthesis in the light and provide the cell with ATP in the light and in the dark. The data presented here serve in identifying changes in protein:protein interactions (PPIs) of Arabidopsis thaliana leaf mitochondria in response to illumination and pave the way towards an understanding how PPIs affect and regulate mitochondrial metabolism.
Project description:The multienzyme glycine cleavage system (GCS) converts glycine and tetrahydrofolate to the one-carbon compound 5,10-methylenetetrahydrofolate, which is of vital importance for most if not all organisms. Photorespiring plant mitochondria contain very high levels of GCS proteins organized as a fragile glycine decarboxylase complex (GDC). The aim of this study is to provide mass spectrometry-based stoichiometric data for the plant leaf GDC and examine whether complex formation could be a general property of the GCS in photosynthesizing organisms. To this end, we investigated the mitochondrial content, the stoichiometry and the isoprotein composition of the Arabidopsis thaliana leaf GDC in comparison with that of Pisum sativum using stable-isotope-labeled peptides (QconCAT and synthetic labeled peptides) and compared the results to label-free, Hi3 peptide quantification.
Project description:Here, we have used digital genomic footprinting to precisely define protein localization for several adipogenic transcription factors at a genome-wide level. In combination with ChIP-seq data, these analyses reveal novel molecular insight into the organization of transcription factors at hotpot regions, which provides a new framework for understanding transcription factor cooperativity on chromatin. Digital genomic footprinting and gene expression in 3T3-L1 pre-adipocytes by high throughput sequencing.
Project description:To evaluate the genome-wide changes in gene translational efficiency during the development of heart failure, we performed transverse aortic constriction(TAC) in male C57BL/6 mice. According to our experience, hypertrophy of the left ventricle was observed 2 weeks after TAC. Cardiac decompensation was observed at 5 weeks. We collected left ventricular tissues at 0, 2, 5 weeks after TAC and then performed ribosome footprinting and sequencing.
Project description:Protein abundance data from S. cerevisiae cells cultivated in synthetic minimal media with a range of metal concentrations (concentrations of Ca, Cu, Fe, K, Mg, Mn, Na and Zn were changed, one at a time)