Structural basis of GTPase-mediated mitochondrial ribosome biogenesis and recycling

• Published in: Nature communications Vol. 12; no. 1; p. 3672
• Main Authors: Hillen, Hauke S., Lavdovskaia, Elena, Nadler, Franziska, Hanitsch, Elisa, Linden, Andreas, Bohnsack, Katherine E., Urlaub, Henning, Richter-Dennerlein, Ricarda
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16-06-2021; Nature Publishing Group; Nature Portfolio
• Subjects: 101/28; 631/337/1910; 631/337/574/1789; 631/45/612/1249; 631/535/1258/1259; 82/83; Assembly; Biosynthesis; Cryoelectron Microscopy; Folding; GTP-Binding Proteins - chemistry; GTP-Binding Proteins - genetics; GTP-Binding Proteins - metabolism; Guanosine triphosphatases; Humanities and Social Sciences; Humans; Intermediates; Maturation; Methyltransferases - chemistry; Methyltransferases - metabolism; Mitochondria; Mitochondrial Ribosomes - chemistry; Mitochondrial Ribosomes - metabolism; Models, Molecular; Molecular machines; Monomeric GTP-Binding Proteins - chemistry; Monomeric GTP-Binding Proteins - metabolism; multidisciplinary; Multiprotein Complexes; Organelle Biogenesis; Peptidyl Transferases - chemistry; Peptidyl Transferases - metabolism; Protein Folding; Quality control; Ribosomal proteins; Ribosome Subunits, Large - chemistry; Ribosome Subunits, Large - metabolism; Ribosomes; RNA, Ribosomal - chemistry; RNA, Ribosomal - metabolism; rRNA; Science; Science (multidisciplinary); Transcription Factors - chemistry; Transcription Factors - metabolism; Yeast
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-23702-y

Ribosome biogenesis requires auxiliary factors to promote folding and assembly of ribosomal proteins and RNA. Particularly, maturation of the peptidyl transferase center (PTC) is mediated by conserved GTPases, but the molecular basis is poorly understood. Here, we define the mechanism of GTPase-driven maturation of the human mitochondrial large ribosomal subunit (mtLSU) using endogenous complex purification, in vitro reconstitution and cryo-EM. Structures of transient native mtLSU assembly intermediates that accumulate in GTPBP6-deficient cells reveal how the biogenesis factors GTPBP5, MTERF4 and NSUN4 facilitate PTC folding. Addition of recombinant GTPBP6 reconstitutes late mtLSU biogenesis in vitro and shows that GTPBP6 triggers a molecular switch and progression to a near-mature PTC state. Additionally, cryo-EM analysis of GTPBP6-treated mature mitochondrial ribosomes reveals the structural basis for the dual-role of GTPBP6 in ribosome biogenesis and recycling. Together, these results provide a framework for understanding step-wise PTC folding as a critical conserved quality control checkpoint.
Maturation of the ribosomal peptidyl transferase center (PTC) is mediated by universally conserved GTPases. Here, cryo-EM structures of mitochondrial ribosomal large subunit assembly intermediates and of mature ribosomes offer insight into the roles of several assembly factors, including GTPBP6’s role in both ribosome biogenesis and recycling.