High-speed atomic force microscopy tracks the dynamic parts of the ribosome

Scheuring, Simon

doi:10.1073/pnas.2024413118

Cited by 3 publications

(1 citation statement)

References 17 publications

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“…In bacteria, lateral stalk binds multiple copies of EF-Tu–aminoacyl–tRNA-GTP ternary complexes to create a high local concentration of translation factors and delivers them to the GTPase-associated center rapidly and accurately ( 14 ). Recently, this suggestion is further supported by high-speed atomic force microscopy, which shows that the P-complex collects archaeal translation factors aEF1α and aEF2 to enrich their local concentration near archaeal ribosomes ( 15 , 16 ). Deletion of both copies of RPL12A and RPL12B genes encoding uL11 in yeast resulting in significant decreases in both cell growth rate and ribosome translation activity ( 17 , 18 ), which could be rescued by transformation with an intact copy of the uL11 gene ( 17 ).…”

Section: Introductionmentioning

confidence: 89%

OUP accepted manuscript

Uchiumi

2022

Nucleic Acids Research

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Eukaryotic uL11 contains a conserved MPPKFDP motif at the N-terminus that is not found in archaeal and bacterial homologs. Here, we determined the solution structure of human uL11 by NMR spectroscopy and characterized its backbone dynamics by 15N–1H relaxation experiments. We showed that these N-terminal residues are unstructured and flexible. Structural comparison with ribosome-bound uL11 suggests that the linker region between the N-terminal domain and C-terminal domain of human uL11 is intrinsically disordered and only becomes structured when bound to the ribosomes. Mutagenesis studies show that the N-terminal conserved MPPKFDP motif is involved in interacting with the P-complex and its extended protuberant domain of uL10 in vitro. Truncation of the MPPKFDP motif also reduced the poly-phenylalanine synthesis in both hybrid ribosome and yeast mutagenesis studies. In addition, G→A/P substitutions to the conserved GPLG motif of helix-1 reduced poly-phenylalanine synthesis to 9–32% in yeast ribosomes. We propose that the flexible N-terminal residues of uL11, which could extend up to ∼25 Å from the N-terminal domain of uL11, can form transient interactions with the uL10 that help to fetch and fix it into a position ready for recruiting the incoming translation factors and facilitate protein synthesis.

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