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“Identifying the SLC25A46 interactome in transgenic mice: cerebellum”

ABSTRACT: SLC25A46 is an outer mitochondrial protein, member of the mitochondrial carrier family (SLC25), whereas mutations result in mitochondrial structure abnormalities leading to a wide spectrum of neurological disorders. SLC25A46 is known to interact with members of the MICOS (Mitochondrial Contact Site) complex involved in mitochondrial cristae organization, and components of the endoplasmic reticulum membrane. In the present work, our aim was to identify the interactome of SLC25A46 in cerebrum, cerebellum, heart and thymus from TgSLC25A46-FLAG transgenic mice through immunoprecipitation with anti-FLAG beads and proteomic analysis with LC-MS/MS.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cerebellum

SUBMITTER: Martina Samiotaki

LAB HEAD: Eleni Douni

PROVIDER: PXD027599 | Pride | 2022-01-11

REPOSITORIES: Pride

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“Identifying the SLC25A46 interactome in transgenic mice: Thymus"

Project description:SLC25A46 is an outer mitochondrial protein, member of the mitochondrial carrier family (SLC25), whereas mutations result in mitochondrial structure abnormalities leading to a wide spectrum of neurological disorders. SLC25A46 is known to interact with members of the MICOS (Mitochondrial Contact Site) complex involved in mitochondrial cristae organization, and components of the endoplasmic reticulum membrane. In the present work, our aim was to identify the interactome of SLC25A46 in cerebrum, cerebellum, heart and thymus from TgSLC25A46-FLAG transgenic mice through immunoprecipitation with anti-FLAG beads and proteomic analysis with LC-MS/MS.

2022-01-11 | PXD027603 | Pride

“Identifying the SLC25A46 interactome in transgenic mice: Heart"

2022-01-11 | PXD027622 | Pride

“Identifying the SLC25A46 interactome in transgenic mice: cerebrum”

Project description:SLC25A46 is an outer mitochondrial protein, member of the mitochondrial carrier family (SLC25), whereas mutations result in mitochondrial structure abnormalities leading to a wide spectrum of neurological disorders. SLC25A46 is known to interact with members of the MICOS (Mitochondrial Contact Site) complex involved in mitochondrial cristae organization, and components of the endoplasmic reticulum membrane. In the present work, our aim was to identify the interactome of SLC25A46 in cerebrum from TgSLC25A46-FLAG transgenic mice through immunoprecipitation with anti-FLAG beads and proteomic analysis with LC-MS/MS.

2022-01-11 | PXD028421 | Pride

Integrative Genomics Positions MKRN1 as a Novel Ribonucleoprotein within the Embryonic Stem Cell Gene Regulatory Network

Project description:For identification of proteins that associate with Makorin1 (MKRN1) in RNA-dependent and RNA-independent manners, we affinity purified FLAG-tagged Makorin1 (MKRN1) from mouse embryonic stem cells constitutively expressing FLAG:MKRN1. Anti-FLAG control immunoprecipitations were performed from a FLAG vectrol control (FLAG:Ctrl) mouse embryonic stem cell line that did not express FLAG:MKRN1. Following FLAG immunoprecipitation, anti-FLAG beads from FLAG:MKRN1 and FLAG:Ctrl immunoprecipitations were split into separate tubes such that half of the beads were digested with 200Ã∞â≈ Ã≠Âµg/mL RNase A while the other half of the beads were undigested. RNase A-digested and undigested immunoprecipitates were subjected to LC-MS/MS analysis. Of the 48 RNA-related proteins previously identified to associate with FLAG:MKRN1, L1TD1, PABPC1, PABPC4, YBX1, IGF2BP1 and UPF1 were found to remain associated with FLAG:MKRN1 in the presence of RNase A.

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NSUN3 and ABH1 modify the wobble position of mt-tRNAMet to expand codon recognition in mitochondrial translation

Project description:Mitochondrial gene expression uses a non-universal genetic code in mammals. Besides reading the conventional AUG codon, mitochondrial (mt-)tRNAMet mediates incorporation of methionine on AUA and AUU codons during translation initiation and on AUA codons during elongation. We show that the RNA methyltransferase NSUN3 localises to mitochondria and interacts with mt-tRNAMet to methylate cytosine 34 (C34) at the wobble position. NSUN3 specifically recognises the anticodon stem loop (ASL) of the tRNA, explaining why a mutation that compromises ASL basepairing leads to disease. We further identify ALKBH1/ABH1 as the dioxygenase responsible for oxidising m5C34 of mt-tRNAMet to generate an f5C34 modification. In vitro codon recognition studies with mitochondrial translation factors reveal preferential utilization of m5C34 mt-tRNAMet in initiation. Depletion of either NSUN3 or ABH1 strongly affects mitochondrial translation in human cells, implying that modifications generated by both enzymes are necessary for mt-tRNAMet function. Together, our data reveal how modifications in mt-tRNAMet are generated by the sequential action of NSUN3 and ABH1, allowing the single mitochondrial tRNAMet to recognise the different codons encoding methionine. HEK293 cell lines expressing His-FLAG-tagged NSUN3 or the His-FLAG tag alone were crosslinked using UV or treated with 5-azacytidine and analysed by CRAC

2016-08-09 | E-GEOD-84664 | biostudies-arrayexpress

Integrative Genomics Positions MKRN1 as a Novel Ribonucleoprotein within the Embryonic Stem Cell Gene Regulatory Network

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2018-10-19 | PXD001191 | Pride

The Slc25a47 locus is a novel determinant of hepatic mitochondrial function implicated in liver fibrosis

Project description:Background & Aims: Transporters of the SLC25 mitochondrial carrier superfamily bridge cytoplasmic and mitochondrial metabolism by channeling metabolites across mitochondrial membranes and are pivotal for metabolic homeostasis. Despite their physiological relevance as gatekeepers of cellular metabolism, most of the SLC25 family members remain uncharacterized. We undertook a comprehensive tissue distribution analysis of all Slc25 family members across metabolic organs and identified SLC25A47 as a liver-specific mitochondrial carrier. Method: We used a murine loss-of-function (LOF) model to unravel the role of this transporter in mitochondrial and hepatic homeostasis. We performed extensive metabolic phenotyping and molecular characterization of Slc25a47hep-/- mice. Results: Slc25a47hep-/- mice displayed a wide variety of metabolic abnormalities, as a result of sustained energy deficiency in the liver originating from impaired mitochondrial respiration in this organ. This mitochondrial phenotype was associated with a robust activation of the mitochondrial stress response in the liver, which in turn, induced the secretion of several mitokines, amongst which FGF21 played a preponderant role in the translation of the effects of the MSR on systemic physiology. To dissect the FGF21-dependent and -independent physiological changes induced by the loss of Slc25a47, we generated a double Slc25a47-Fgf21 LOF mouse model, and demonstrated that several aspects of the hypermetabolic state was entirely driven by hepatic secretion of FGF21. On the other hand, the metabolic fuel inflexibility induced by loss of Slc25a47 could not be rescued by genetical removal of Fgf21. Conclusion: Collectively, our data place SLC25A47 at the center of mitochondrial homeostasis, which upon dysfunction triggers robust liver-specific and systemic adaptive stress responses.

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Transcription profiling of human fetal cerebellum, heart, cerebrum, and cerebrum derived astrocyte cell line from trisomy 21 and trisomy 13 samples

Project description:Gene expression was measured in trisomy 21 and trisomy 13 human fetal samples. For TS21, regions assayed were cerebrum, cerebellum, heart, and cerebrum-derived astrocyte cell lines.

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The MICOS-MIB complex member PERM1 connects mitochondria with the sarcolemma via ankyrin B

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2021-11-16 | PXD025745 | Pride

A multiple reporter protein tolerant mouse model

Project description:We report the generation of a novel mouse strain that is immunologically tolerant to exogenously introduced reporter proteins ('Tol' strain). As a control, we test whether any transcriptomic changes can be observed between the Tol strain and wild-type littermate. This is tested for thymus, cerebrum and cerebellum.

2020-04-12 | GSE147757 | GEO

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