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Cardiac mitochondrial function depends on BUD23 mediated ribosome programming

Elife. 2020; 
Baxter M, , Voronkov M, , Poolman T, , Galli G, Pinali C, Goosey L, Knight A, Krakowiak K, Maidstone R, , Iqbal M, Zi M, Prehar S, Cartwright EJ, Gibbs J, Matthews LC, Adamson AD, Humphreys NE, Rebelo-Guiomar P, Minczuk M, Bechtold DA, Loudon A, Ray D,
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Recombinant Proteins A double stranded DNA (dsDNA) repair template was designed to (i) incorporate loxP sites, with unique restriction sites for screening purposes, 300 bp upstream and 200 bp downstream of exon seven respectively and (ii) integrate silent shield mutations for all three sgRNA to prevent cutting of the repair template and (iii) contain 1000 bp 5’ and 3’ homology arms to the target region. The above template was synthesised in a pUC57 vector (Genscript, US), the linear 5’homology-loxP-exon7-loxP-3’homology fragment excised from the vector by restriction digest, gel extracted (Bioline) and further cleaned prior to microinjection (Monarch PCR purification kit, NEB). Get A Quote

摘要

Efficient mitochondrial function is required in tissues with high energy demand such as the heart, and mitochondrial dysfunction is associated with cardiovascular disease. Expression of mitochondrial proteins is tightly regulated in response to internal and external stimuli. Here we identify a novel mechanism regulating mitochondrial content and function, through BUD23-dependent ribosome generation. BUD23 was required for ribosome maturation, normal 18S/28S stoichiometry and modulated the translation of mitochondrial transcripts in human A549 cells. Deletion of Bud23 in murine cardiomyocytes reduced mitochondrial content and function, leading to severe cardiomyopathy and death. We discovered that BUD23 selectiv... More

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