Mutations in the leader region of ribosomal RNA operons cause structurally defective 30 S ribosomes as revealed by in vivo structural probing

Journal of Molecular Biology
M Balzer, R Wagner

Abstract

The biogenesis of functional ribosomes is regulated in a very complex manner, involving different proteins and RNA molecules. RNAs are not only essential components of both ribosomal subunits but also transiently interacting factors during particle formation. In eukaryotes snoRNAs act as molecular chaperones to assist maturation, modification and assembly. In a very similar way highly conserved leader sequences of bacterial rRNA operons are involved in the correct formation of 30 S ribosomal subunits. Certain mutations in the rRNA leader region cause severe growth defects due to malfunction of ribosomes which are assembled from such transcription units. To understand how the leader sequences act to facilitate the formation of the correct 30 S subunits we performed in vivo chemical probing to assess structural differences between ribosomes assembled either from rRNA transcribed from wild-type operons or from operons which contain mutations in the rRNA leader region. Cells transformed with plasmids containing the respective rRNA operons were reacted with dimethylsulphate (DMS). Ribosomes were isolated by sucrose gradient centrifugation and modified nucleotides within the 16 S rRNA were identified by primer extension reaction. Str...Continue Reading

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Citations

Jul 7, 2011·The Journal of Biological Chemistry·Lucyna LubkowskaNatalia Komissarova
Aug 10, 2004·BMC Molecular Biology·Irmtraud M Meyer, István Miklós
Oct 18, 2013·RNA·Daniel LaiIrmtraud M Meyer
May 21, 2003·RNA·Susan L Heilman-Miller, Sarah A Woodson
Sep 10, 2014·Nature Structural & Molecular Biology·Neha Gupta, Gloria M Culver
Nov 4, 2010·RNA Biology·Georgeta Zemora, Christina Waldsich
Sep 1, 2008·EcoSal Plus·Gloria M Culver, Narayanaswamy Kirthi

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