Structure of the two most C-terminal RNA recognition motifs of PTB using segmental isotope labeling

Francesca Vitali, Anke Henning, Florian C. Oberstrass, Yann Hargous, Sigrid D. Auweter, Michèle Erat, Frédéric H.T. Allain

Research output: Contribution to journalArticle

71 Scopus citations

Abstract

The polypyrimidine tract binding protein (PTB) is a 58 kDa protein involved in many aspects of RNA metabolism. In this study, we focused our attention on the structure of the two C-terminal RNA recognition motifs (RRM3 and RRM4) of PTB. In a previous study, it was found that the two RRMs are independent in the free state. We recently determined the structure of the same fragment in complex with RNA and found that the two RRMs interact extensively. This difference made us re-evaluate in detail the free protein structure and in particular the interdomain interface. We used a combination of NMR spectroscopy and segmental isotopic labeling to unambiguously study and characterize the interdomain interactions. An improved segmental isotopic labeling protocol was used, enabling us to unambiguously identify 130 interdomain NOEs between the two RRMs and to calculate a very precise structure. The structure reveals a large interdomain interface, resulting in a very unusual positioning of the two RRM domains relative to one another.

Original languageEnglish (US)
Pages (from-to)150-162
Number of pages13
JournalEMBO Journal
Volume25
Issue number1
DOIs
StatePublished - Jan 11 2006
Externally publishedYes

Keywords

  • Isotopic labeling
  • NMR
  • Protein ligation
  • RRM/RBD/RNP
  • Segmental labeling

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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    Vitali, F., Henning, A., Oberstrass, F. C., Hargous, Y., Auweter, S. D., Erat, M., & Allain, F. H. T. (2006). Structure of the two most C-terminal RNA recognition motifs of PTB using segmental isotope labeling. EMBO Journal, 25(1), 150-162. https://doi.org/10.1038/sj.emboj.7600911