Counteraction by MutT Protein of Transcriptional Errors Caused by Oxidative Damage

Taddéi, François; Hayakawa, Hiroshi; Bouton, Marie‐Florence; Cirinesi, Anne Marie; Matić, Ivan; Sekiguchi, Mutsuo; Radman, Miroslav

doi:10.1126/science.278.5335.128

Cited by 263 publications

(267 citation statements)

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“…Thus, 8-OH-Gua could be formed in cellular RNA via the incorporation of 8-OH-GTP. Because this oxidized ribonucleotide has been suggested to be involved in transcriptional and translational errors [28], the oxidation of GTP would perturb the accurate expression of genetic information.…”

Section: Discussionmentioning

confidence: 99%

“…8-OH-GTP was previously used in in vitro transcription reactions conducted by RNA pols from phage T7, E. coli, and calf thymus [28,56,57]. Poly(dA-dT), E. coli and calf thymus DNAs, and linearized plasmid DNA were employed as templates in the previous studies, and thus a detailed analysis of 8-OH-GTP incorporation was difficult.…”

Section: Discussionmentioning

confidence: 99%

“…Because the pool size of ribonucleotides is hundreds of times larger than that of 2'-deoxyribonucleotides [26,27], oxidized RNA precursors may be more abundant than oxidized DNA precursors. Previously, Taddei 8-oxo-7,8-dihydroguanosine 5'-triphosphate), was incorporated into RNA [28] and concluded that 8-OH-GTP induced translational errors. The findings that the E. coli MutT, and mammalian MTH1 (NUDT1) and NUDT5 proteins catalyze the hydrolysis of oxidized RNA precursors suggest the importance of the hydrolytic elimination of oxidized ribonucleotides [28][29][30].…”

mentioning

confidence: 99%

“…Previously, Taddei 8-oxo-7,8-dihydroguanosine 5'-triphosphate), was incorporated into RNA [28] and concluded that 8-OH-GTP induced translational errors. The findings that the E. coli MutT, and mammalian MTH1 (NUDT1) and NUDT5 proteins catalyze the hydrolysis of oxidized RNA precursors suggest the importance of the hydrolytic elimination of oxidized ribonucleotides [28][29][30]. Moreover, disruption of the transcription factor S-II, which enhances the excision of ribonucleotides misincorporated by RNA polymerase (pol) II in vitro, confers sensitivity to oxidants together with low transcriptional fidelity in yeast, and antioxidant treatment relieves this low transcriptional fidelity [31].…”

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confidence: 99%

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Incorporation of 8-hydroxyguanosine (8-oxo-7,8-dihydroguanosine) 5′-triphosphate by bacterial and human RNA polymerases

Kamiya

Suzuki

Yamaguchi

et al. 2009

Free Radical Biology and Medicine

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Oxidized RNA precursors formed in the nucleotide pool may be incorporated into RNA. In this study, the incorporation of is one of the most abundant ([5] and references therein). The highly reactive hydroxy radical and guanine-selective singlet oxygen are thought to be involved in the formation of 8-OH-Gua [6]. 8-OH-Gua can pair with both cytosine and adenine [7][8][9][10]. The oxidized nucleobase in DNA seems to be an important source of mutations, and it induces G:CT:A transversions in living cells [11][12][13][14][15][16][17][18][19]. The oxidation of dGTP in the nucleotide pool by reactive oxygen species is another significant source in the mutation process. 8-Hydroxy-2'-deoxyguanosine 5'-triphosphate In this study, we analyzed the incorporation of 8-OH-GTP by the E. coli and human RNA pols in vitro. We found that the oxidized ribonucleotide was incorporated into RNA opposite C and A in the template DNA. This result suggests that 8-OH-GTP is incorporated into RNA, resulting in transcriptional and translational errors. In contrast,another oxidized ribonucleotide, 2-hydroxyadenosine 5'-triphosphate (2-OH-ATP) [29], was incorporated "correctly" opposite T. Materials and methods Materials 6E. coli RNA polymerase was purchased from Sigma-Aldrich (St.Louis, MO, USA). Human RNA pol II was purified from HeLa cells expressing Flag-and His-tagged human Rpb3 (the third subunit of RNA pol II), as described previously [32]. Ribonuclease inhibitor was from Takara (Kusatsu, Japan). 8-OH-GTP and 2-OH-ATP were prepared by the oxidation of GTP and ATP, respectively, as described previously [29]. The purities of the 8-OH-GTP and 2-OH-ATP were both estimated to be more than 99%, as determined by an HPLC analysis (data not shown). The unmodified ribonucleoside 5'-triphosphates were from GE HealthcareBio-Sciences (Piscataway, NJ, USA). The Alexa Fluor 647 (Alexa 647)-modified oligoribonucleotide and the unmodified oligodeoxyribonucleotides (Table 1) were purchased from Japan Bio Services (Asaka, Japan) and Invitrogen Japan (Tokyo, Japan), respectively, in purified forms. In vitro RNA synthesisAn RNA primer-extension assay, in which the elongation complexes were assembled on nucleic acid scaffolds [33][34][35][36][37][38] Hepes-NaOH (pH 7.9), 5 mM MgCl 2 , 0.5 mM EDTA, 0.5 mM dithiothreitol, 0.5 U/µl ribonuclease inhibitor, 500 µM NTP and the enzyme at 30°C. In both reactions, the RNA/DNA hybrid was incubated with RNA pol for 5 min to assemble the elongation complex (preincubation), and then the substrate ribonucleotide was added to start the reactions.Reactions were stopped by the addition of a two-fold volume of termination solution (90% formamide, 50 mM EDTA). Samples were heated at 98°C for 5 min and chilled on ice, and were then applied to an 8 M urea, 20% polyacrylamide gel. Fluorograms were obtained with a Ε. coli RNA pol were carried out at 37°C for two to ten min. Results Incorporation of 8-OH-GTP by E. coli RNA polymeraseTo monitor nucleotide incorporation, we utilized a primer-extension assay, in which the elon...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Incorporation of 8-hydroxyguanosine (8-oxo-7,8-dihydroguanosine) 5′-triphosphate by bacterial and human RNA polymerases

Kamiya

Suzuki

Yamaguchi

et al. 2009

Free Radical Biology and Medicine

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“…In the mutT strain, a protein containing an altered sequence was produced with higher frequency than that expected from the mutation frequency at the DNA level [13]. The authors of that paper concluded that an oxidized form of GTP, 8-OH-GTP (also known as 8-oxo-7,8-dihydroguanosine 5'-triphosphate), was misincorporated opposite A by RNA pol and induced translational errors, because the MutT protein hydrolyzes 8-OH-GTP to the monophosphate [13]. The findings that some MutT-type enzymes including human MTH1 protein catalyze the hydrolysis of oxidized RNA precursors suggest the importance of the hydrolytic elimination of oxidized ribonucleotides [ref.…”

Section: Introductionmentioning

confidence: 96%

Effects of 8-hydroxy-GTP and 2-hydroxy-ATP on in vitro transcription

Kamiya

Suzuki

Kawai

et al. 2007

Free Radical Biology and Medicine

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Factors contributing to the outcome of oxidative damage to nucleic acids

2004

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Oxidative damage to DNA appears to be a factor in cancer, yet explanations for why highly elevated levels of such lesions do not always result in cancer remain elusive. Much of the genome is non-coding and lesions in these regions might be expected to have little biological effect, an inference supported by observations that there is preferential repair of coding sequences. RNA has an important coding function in protein synthesis, and yet the consequences of RNA oxidation are largely unknown. Some non-coding nucleic acid is functional, e.g. promoters, and damage to these sequences may well have biological consequences. Similarly, oxidative damage to DNA may promote microsatellite instability, inhibit methylation and accelerate telomere shortening. DNA repair appears pivotal to the maintenance of genome integrity, and genetic alterations in repair capacity, due to single nucleotide polymorphisms or mutation, may account for inter-individual differences in cancer susceptibility. This review will survey these aspects of oxidative damage to nucleic acids and their implication for disease.

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Counteraction by MutT Protein of Transcriptional Errors Caused by Oxidative Damage

Cited by 263 publications

References 25 publications

Incorporation of 8-hydroxyguanosine (8-oxo-7,8-dihydroguanosine) 5′-triphosphate by bacterial and human RNA polymerases

Incorporation of 8-hydroxyguanosine (8-oxo-7,8-dihydroguanosine) 5′-triphosphate by bacterial and human RNA polymerases

Effects of 8-hydroxy-GTP and 2-hydroxy-ATP on in vitro transcription

Factors contributing to the outcome of oxidative damage to nucleic acids

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