PMID: 9917068Jan 23, 1999

Identification of the ribosome binding sites of translation initiation factor IF3 by multidimensional heteronuclear NMR spectroscopy

RNA
M SetteR Boelens

Abstract

Titrations of Escherichia coli translation initiation factor IF3, isotopically labeled with 15N, with 30S ribosomal subunits were followed by NMR by recording two-dimensional (15N,1H)-HSQC spectra. In the titrations, intensity changes are observed for cross peaks belonging to amides of individual amino acids. At low concentrations of ribosomal subunits, only resonances belonging to amino acids of the C-domain of IF3 are affected, whereas all those attributed to the N-domain are still visible. Upon addition of a larger amount of 30S subunits cross peaks belonging to residues of the N-terminal domain of the protein are also selectively affected. Our results demonstrate that the two domains of IF3 are functionally independent, each interacting with a different affinity with the ribosomal subunits, thus allowing the identification of the individual residues of the two domains involved in this interaction. Overall, the C-domain interacts with the 30S subunits primarily through some of its loops and alpha-helices and the residues involved in ribosome binding are distributed rather symmetrically over a fairly large surface of the domain, while the N-domain interacts mainly via a small number of residues distributed asymmetrically in t...Continue Reading

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Citations

Aug 6, 2003·Journal of Molecular Biology·D PetrelliA La Teana
May 6, 2003·Protein Expression and Purification·Alexandra WolfrumNorbert Grillenbeck
Feb 26, 2009·Biochemistry·Brooke E Christian, Linda L Spremulli
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May 24, 2003·Cold Spring Harbor Symposia on Quantitative Biology·A BashanAda Yonath
Mar 10, 2005·Microbiology and Molecular Biology Reviews : MMBR·Brian Søgaard LaursenHans Uffe Sperling-Petersen
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Aug 12, 2015·Cellular and Molecular Life Sciences : CMLS·Claudio O Gualerzi, Cynthia L Pon
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Apr 21, 2012·Critical Reviews in Biochemistry and Molecular Biology·Pohl Milón, Marina V Rodnina
Dec 15, 2010·The Journal of Biological Chemistry·Lei WeiKonstantin Pervushin

Related Concepts

Bacterial Proteins
Alkalescens-Dispar Group
In Vivo NMR Spectroscopy
Peptide Initiation Factors
Protein Conformation
Ribonucleoproteins
Ribosomes
RNA-Binding Proteins
Protein Conformation, beta-Strand
Eukaryotic Initiation Factor-3, Subunit 7

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