Dataset Information

Label-free shotgun proteomics of M. hyopneumoniae and M. feriruminatoris

ABSTRACT: Mycoplasmas are wall-less parasitic bacteria possessing extremely small genomes. Despite this, growth rates differ among mycoplasma species with doubling times ranging from only 0.5h to 16h. Here, we analyzed the whole proteomes of M. hyopneumoniae and M. feriruminatoris and performed a comparative study in slow and fast growing mycoplasmas.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mycoplasma Feriruminatoris Mycoplasma Hyopneumoniae (strain 232)

SUBMITTER: Marc Weber

LAB HEAD: Luis Serrano

PROVIDER: PXD025422 | Pride | 2021-08-23

REPOSITORIES: Pride

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Publications

Widespread ribosome stalling in a genome-reduced bacterium and the need for translational quality control.

Burgos Raul R Weber Marc M Gallo Carolina C Lluch-Senar Maria M Serrano Luis L

iScience 20210816 9

Trans-translation is a ubiquitous bacterial mechanism of ribosome rescue mediated by a transfer-messenger RNA (tmRNA) that adds a degradation tag to the truncated nascent polypeptide. Here, we characterize this quality control system in a genome-reduced bacterium, <i>Mycoplasma pneumoniae</i> (MPN), and perform a comparative analysis of protein quality control components in slow and fast-growing prokaryotes. We show <i>in vivo</i> that in MPN the sole quality control cytoplasmic protease (Lon) d ...[more]

PMID: 34485867

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Project description:DHX15 is an ATP-dependent RNA helicase involved in pre-mRNA splicing. We have recently reported that DHX15 is a downstream substrate for Akt1, which plays a significant role in vascular biology. Therefore, we aimed to explore the regulatory function of DHX15 over the vasculature, and the endothelial cell biology in different contexts: development, metabolism, ischemia and tumor growth. Methods: Deficient DHX15 mice and zebrafish were generated using transcription activator-like effector nuclease (TALEN) and Crispr/cas9 gene edition. Lymphatic functionality was evaluated by lymphangiography and magnetic resonance imaging. Mouse-induced tumor and metastasis model were generated by injection of syngeneic LLC1 tumor cells. DHX15 gene silencing in mouse liver endothelial cells (LEC) was performed by the lentiviral transduction of shRNA (Dharmacon). The changes in the transcriptome and the proteome resulting from the shRNA transduction were investigated by RNAseq and mass spectrometry, respectively. Results: Homozygous DHX15 gene deficiency was lethal in mouse and zebrafish embryos. DHX15-/- zebrafish also showed an undeveloped parachordal line, which leads to the formation of lymphatic structures in the trunk during the development. DHX15+/- mice and zebrafish were viable, although DHX15+/- gene deficiency triggered lower vascular network density and impaired lymphatic function postnatally in mice. Whole transcriptome and proteome analysis of DHX15 silenced LEC revealed differential expression of enzymes involved in the glycolysis and the gluconeogenesis pathways. The functional validation of these results demonstrated an uncoupling of the glycolysis with the oxidation of pyruvate into the mitochondria and lower activity of the Complex I in the mitochondrial membrane, resulting in lower cellular ATP production. Noteworthy, heterozygous DHX15 deficiency partially inhibited primary tumor growth and reduced lung metastasis after injection of syngeneic LLC1 tumor cells, compared to wild-type mice

Dataset Information

Label-free shotgun proteomics of M. hyopneumoniae and M. feriruminatoris

Publications

Widespread ribosome stalling in a genome-reduced bacterium and the need for translational quality control.

Similar Datasets

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