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| Title | The late stages of yeast mitoribosome large subunit biogenesis. |
|---|---|
| Journal, issue, pages | Biochim Biophys Acta Mol Cell Res, Vol. 1872, Issue 8, Page 120051, Year 2025 |
| Publish date | Aug 25, 2025 |
 Authors | Sorbhi Rathore / Julian Conrad / Dasmanthie De Silva / Alberto Ferrari / Danielle Bouquio / Hyung-Jun Kim / Roger Salvatori / Andreas Linden / Olexandr Dybkov / Henning Urlaub / Martin Ott / Antoni Barrientos /    |
| PubMed Abstract | The Saccharomyces cerevisiae mitoribosome synthesizes eight mitochondrial DNA-encoded proteins essential for oxidative phosphorylation. Mitoribosome large subunit (mtLSU) biogenesis involves the ...The Saccharomyces cerevisiae mitoribosome synthesizes eight mitochondrial DNA-encoded proteins essential for oxidative phosphorylation. Mitoribosome large subunit (mtLSU) biogenesis involves the conserved DEAD-box helicase Mrh4 and the GTPases Mtg1/GTPBP7 and Mtg2/GTPBP5. Here, we have employed genetic, biochemical, in vitro reconstitution, and cryo-EM approaches to elucidate their hierarchical action during the late stages of mtLSU assembly. We show that Mrh4-mediated bL33m incorporation precedes Mtg1 recruitment to the 21S rRNA. Cryo-EM structures of mitoribosome assembly intermediates accumulating in the absence of Mtg1 or uL16m reveal that Mtg1 restructures the 21S rRNA H73-75 and H93 domains to their mature fold. This subsequently allows the structuring of neighboring peptidyl transfer center region helices and the incorporation of uL6m, uL16m, bL35m, and bL36m during late mtLSU maturation. Unexpectedly, monosomes containing immature mtLSU assemble in Mrh4-, bL33m-, uL16m-, Mtg1-, and Mtg2-depleted mitochondria, at levels that increase with the maturation state of the mtLSU particle. Our data have shed light on the rRNA folding events and the structuring of the MRPs that occur during the late stages of assembly. They have provided insight into the roles of assembly factors Mrh4, Mtg1, and Mtg2 during the process and revealed evolutionarily conserved mechanisms underlying mitochondrial ribosome assembly. |
 External links | Biochim Biophys Acta Mol Cell Res / PubMed:40865570 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 3.4 - 6.0 Å |
| Structure data |  EMDB-16559: Structure of the yeast delta mtg1 mitochondrial ribosome assembly intermediate - State1  EMDB-16560: Structure of the yeast delta mtg1 mitochondrial ribosome assembly intermediate - State 2  EMDB-16561: Structure of the yeast delta mtg1 mitochondrial ribosome - State 3  EMDB-16562: Structure of the yeast delta uL16m mitochondrial ribosome assembly intermediate - State 1  EMDB-16590: Structure of the yeast delta uL16m mitochondrial ribosome - State 2 |
| Source |
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Saccharomyces cerevisiae W303 (yeast)