Detection and discovery of RNA modifications using microarrays

Nucleic Acids Res. 2005 Jan 7;33(1):e2. doi: 10.1093/nar/gni002.

Abstract

Using a microarray that tiles all known yeast non-coding RNAs, we compared RNA from wild-type cells with RNA from mutants encoding known and putative RNA modifying enzymes. We show that at least five types of RNA modification (dihydrouridine, m1G, m2(2)G, m1A and m6(2)A) catalyzed by 10 different enzymes (Trm1p, Trm5, Trm10p, Dus1p-Dus4p, Dim1p, Gcd10p and Gcd14p) can be detected by virtue of differential hybridization to oligonucleotides on the array that are complementary to the modified sites. Using this approach, we identified a previously undetected m1A modification in GlnCTG tRNA, the formation of which is catalyzed by the Gcd10/Gcd14 complex. complex.

Publication types

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

MeSH terms

  • Mutation
  • Oligonucleotide Array Sequence Analysis / methods*
  • RNA Processing, Post-Transcriptional*
  • RNA, Fungal / chemistry
  • RNA, Fungal / metabolism
  • RNA, Transfer / chemistry
  • RNA, Transfer / metabolism
  • RNA, Untranslated / chemistry
  • RNA, Untranslated / metabolism*
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*

Substances

  • RNA, Fungal
  • RNA, Untranslated
  • RNA, Transfer