Characterization of a Trypanosoma brucei Alkb homolog capable of repairing alkylated DNA

A Role for Saccharomyces cerevisiae Tpa1 Protein in Direct Alkylation Repair

“…2D). This is in good agreement with the characteristic absorption peak of chromophores reported by others (33,45,46). However, the Tpa1-mutant, which lacks active site coordination of iron and 2OG, did not produce any spectrum associated with chromophore formation (Fig.…”

“…Although Tpa1 has been characterized before (24 -26), this spectroscopic characteristic was not known in Tpa1. We added excess Fe(II) and 2OG to recombinant Tpa1 and observed an UV-visible band with peak absorption at 530 nm for the native protein, as described previously (33,45,46). This is the first Geneticin 80mg/ml WT tpa1D mag1D tpa1D mag1D stp22D WT tpa1D mag1D tpa1D mag1D stp22D 6-aza-uracil 100mg/ml YPD FIGURE 7.…”

“…Members of the Fe(II)/2OG-dependent dioxygenases forms chromophores associated with a metal-to-ligand charge transfer transition. When we monitored purified Tpa1 by UVvisible spectroscopy under the optimum conditions mentioned before (45), it generated the characteristic absorption peak at 530 nm when both metal and cofactor were present (Fig. 2D).…”

Nucleic Acid Oxidation in DNA Damage Repair and Epigenetics

“…Bioinformatics and functional analyses reveal that AlkB homologues are widely expressed in many organisms, from bacteria to humans, and carry out diverse biological functions. 42 , 95 − 103 The majority of bacterial AlkB homologues are DNA-repair proteins. 100 , 103 Two AlkB homologues have been found in the genome of fission yeast S. pombe .…”

The DNA damage response is developmentally regulated in the African trypanosome

“…Genetic and biochemical studies clearly demonstrate the action of Nucleotide Excision Repair (NER) [[27], [28], [29]], Base Excision Repair (BER) [[30], [31], [32], [33], [34], [35], [36], [37], [38]] and Mismatch Repair (MMR) in T. brucei [[39], [40], [41], [42], [43], [44]]. In contrast, further T. brucei DNA repair pathways are less well explored, including Direct Reversion Repair (DR) [45], Interstrand Crosslink Repair (ICL repair) [29,46] and Translesion Synthesis (TLS) [47]. In all cases, the potential that trypanosomatid repair pathways might differ from counterpart pathways in model eukaryotes, reflecting parasite-specific biology, is relatively poorly understood.…”