Pyrite footprinting of RNA

Jörg C. Schlatterer, Matthew S. Wieder, Christopher D. Jones, Lois Pollack, Michael D. Brenowitz

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

In RNA, function follows form. Mapping the surface of RNA molecules with chemical and enzymatic probes has revealed invaluable information about structure and folding. Hydroxyl radicals (OH) map the surface of nucleic acids by cutting the backbone where it is accessible to solvent. Recent studies showed that a microfluidic chip containing pyrite (FeS2) can produce sufficient OH to footprint DNA. The 49-nt Diels-Alder RNA enzyme catalyzes the C-C bond formation between a diene and a dienophile. A crystal structure, molecular dynamics simulation and atomic mutagenesis studies suggest that nucleotides of an asymmetric bulge participate in the dynamic architecture of the ribozyme's active center. Of note is that residue U42 directly interacts with the product in the crystallized RNA/product complex. Here, we use powdered pyrite held in a commercially available cartridge to footprint the Diels-Alderase ribozyme with single nucleotide resolution. Residues C39 to U42 are more reactive to OH than predicted by the solvent accessibility calculated from the crystal structure suggesting that this loop is dynamic in solution. The loop's flexibility may contribute to substrate recruitment and product release. Our implementation of pyrite-mediated OH footprinting is a readily accessible approach to gleaning information about the architecture of small RNA molecules.

Original languageEnglish (US)
Pages (from-to)374-378
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume425
Issue number2
DOIs
StatePublished - Aug 24 2012

Fingerprint

RNA
Catalytic RNA
DNA Footprinting
Nucleotides
Crystal structure
Mutagenesis
Molecules
Microfluidics
Molecular Dynamics Simulation
Hydroxyl Radical
Nucleic Acids
Molecular dynamics
pyrite
DNA
Computer simulation
Substrates
Enzymes

Keywords

  • Footprinting
  • Hydroxyl radical
  • Pyrite
  • RNA

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Pyrite footprinting of RNA. / Schlatterer, Jörg C.; Wieder, Matthew S.; Jones, Christopher D.; Pollack, Lois; Brenowitz, Michael D.

In: Biochemical and Biophysical Research Communications, Vol. 425, No. 2, 24.08.2012, p. 374-378.

Research output: Contribution to journalArticle

Schlatterer, JC, Wieder, MS, Jones, CD, Pollack, L & Brenowitz, MD 2012, 'Pyrite footprinting of RNA', Biochemical and Biophysical Research Communications, vol. 425, no. 2, pp. 374-378. https://doi.org/10.1016/j.bbrc.2012.07.100
Schlatterer, Jörg C. ; Wieder, Matthew S. ; Jones, Christopher D. ; Pollack, Lois ; Brenowitz, Michael D. / Pyrite footprinting of RNA. In: Biochemical and Biophysical Research Communications. 2012 ; Vol. 425, No. 2. pp. 374-378.
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