SNAP output for PDB entry 7v2m [SNAP web server]

PDB-id

7v2m; SNAP-derived features in text and JSON formats; DNAproDB

Class

ribosome

Method

cryo-EM (3.4 Å)

Summary

T.thermophilus 30s ribosome with ksga, class k1k4

Reference

Singh J, Raina R, Vinothkumar KR, Anand R (2022): "Decoding the Mechanism of Specific RNA Targeting by Ribosomal Methyltransferases." Acs Chem.Biol., 17, 829-839. doi: 10.1021/acschembio.1c00732.

Abstract

Methylation of specific nucleotides is integral for ribosomal biogenesis and also serves as a common mechanism to confer antibiotic resistance by pathogenic bacteria. Here, by determining the high-resolution structure of the 30S-KsgA complex by cryo-electron microscopy, a state was captured, where KsgA juxtaposes between helices h44 and h45 of the 30S ribosome, separating them, thereby enabling remodeling of the surrounded rRNA and allowing the cognate site to enter the methylation pocket. With the structure as a guide, several mutant versions of the ribosomes, where interacting bases in the catalytic helix h45 and surrounding helices h44, h24, and h27, were mutated and evaluated for their methylation efficiency revealing factors that direct the enzyme to its cognate site with high fidelity. The biochemical studies show that the three-dimensional environment of the ribosome enables the interaction of select loop regions in KsgA with the ribosome helices paramount to maintain selectivity.