Joint X-ray and NMR refinement of the yeast L30e-mRNA complex

Structure. 2004 Jul;12(7):1165-76. doi: 10.1016/j.str.2004.04.023.

Abstract

L30e, a Saccharomyces cervisiae ribosomal protein, regulates its own expression by binding to a purine-rich asymmetric internal loop located in both its pre-mRNA and mature mRNA. A crystal structure of an MBP-L30e fusion protein in complex with an RNA containing the pre-mRNA regulatory site was solved at 3.24 A. Interestingly, the structure of the RNA differed from that observed in a previously determined NMR structure of the complex. Analysis of the NMR data led to the identification of a single imino proton resonance in the internal loop that had been incorrectly assigned and was principally responsible for the erroneous RNA structure. A structure refinement was performed using both the X-ray diffraction data and the NMR-derived distance and angle restraints. The joint NMR and X-ray refinement resulted in improved stereochemistry and lower crystallographic R factors. The RNA internal loop of the MBP-L30e-mRNA complex adopts the canonical K-turn fold.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Crystallization
  • Crystallography, X-Ray
  • Magnetic Resonance Spectroscopy*
  • Models, Molecular
  • Molecular Weight
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • RNA, Messenger / chemistry*
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / metabolism
  • Ribosomal Proteins / chemistry*
  • Ribosomal Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism

Substances

  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Ribosomal Proteins
  • Saccharomyces cerevisiae Proteins
  • ribosomal protein L30

Associated data

  • PDB/1T0K