Isolation of a site-specifically modified RNA from an unmodified transcript.
Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-specifically modified RNAs away from unmodified transcripts. Using the m1G37 and m1A58 methyl modifications of tRNA as two examples, we demonstrate that non-pairing base modifications protect RNAs against the DNA-directed RNase H cleavage. This provide a new approach to obtain homogeneous RNAs with site-specific base modifications that are suitable for biochemical and functional studies.
Department of Biochemistry and Molecular Biology, Thomas Jefferson University 233 South 10th Street, BLSB 220, Philadelphia, PA 19107-5719, USA. Ya-Ming.Hou@jefferson.edu
Ya-Ming
YM
Hou
Zhi
Z
Li
Howard
H
Gamper
D001483:Base Sequence; D019175:DNA Methylation; D017017:Methanococcus / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry; D012323:RNA Processing, Post-Transcriptional; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism
D009841:Oligonucleotides; D012343:RNA, Transfer; D016914:Ribonuclease H