Article ; Online: Methylation damage to RNA induced in vivo in Escherichia coli is repaired by endogenous AlkB as part of the adaptive response.
2013 Volume 12, Issue 3, Page(s) 188–195
Abstract: Cytotoxic 1-methyladenine (1-meA) and 3-methylcytosine (3-meC) lesions induced in DNA and RNA in vitro and in pre-damaged DNA and RNA bacteriophages in vivo are repaired by the Escherichia coli (E. coli) protein AlkB and a human homolog, ALKBH3. However, ...
| Abstract | Cytotoxic 1-methyladenine (1-meA) and 3-methylcytosine (3-meC) lesions induced in DNA and RNA in vitro and in pre-damaged DNA and RNA bacteriophages in vivo are repaired by the Escherichia coli (E. coli) protein AlkB and a human homolog, ALKBH3. However, it is not known whether endogenous RNA is repaired in vivo by repair proteins present at physiological concentrations. The concept of RNA repair as a biologically relevant process has therefore remained elusive. Here, we demonstrate AlkB-mediated repair of endogenous RNA in vivo by measuring differences in lesion-accumulation in two independent AlkB-proficient and deficient E. coli strains during exposure to methyl methanesulfonate (MMS). Repair was observed both in AlkB-overproducing strains and in the wild-type strains after AlkB induction. RNA repair appeared to be highest in RNA species below 200 nucleotides in size, mainly comprising tRNAs. Strikingly, at least 10-fold more lesions were repaired in RNA than in DNA. This may be a consequence of some 30-fold higher levels of aberrant methylation in RNA than in DNA after exposure to MMS. A high primary kinetic isotope effect (>10) was measured using a deuterated methylated RNA substrate, D3-1me(rA), demonstrating that it is the catalytic step, and not the search step that is rate-limiting. Our results demonstrate that RNA repair by AlkB takes place in endogenous RNA as part of an adaptive response in wild-type E. coli cells. |
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| MeSH term(s) | Adaptation, Physiological/genetics ; Alkylating Agents/pharmacology ; DNA Repair ; DNA, Bacterial/genetics ; Enzyme Induction ; Escherichia coli/enzymology ; Escherichia coli/genetics ; Escherichia coli Proteins/physiology ; Kinetics ; Methyl Methanesulfonate/pharmacology ; Methylation ; Mixed Function Oxygenases/physiology ; RNA, Bacterial/genetics ; RNA, Bacterial/metabolism |
| Chemical Substances | Alkylating Agents ; DNA, Bacterial ; Escherichia coli Proteins ; RNA, Bacterial ; Methyl Methanesulfonate (AT5C31J09G) ; Mixed Function Oxygenases (EC 1.-) ; AlkB protein, E coli (EC 1.14.11.-) |
| Language | English |
| Publishing date | 2013-03-01 |
| Publishing country | Netherlands |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 2071608-4 |
| ISSN | 1568-7856 ; 1568-7864 |
| ISSN (online) | 1568-7856 |
| ISSN | 1568-7864 |
| DOI | 10.1016/j.dnarep.2012.11.010 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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