The AlkB protein is a repair enzyme that uses an α-ketoglutarate/Fe(II)-dependent mechanism to repair alkyl DNA adducts. AlkB has been reported to repair highly susceptible substrates, such as 1-methyladenine and 3-methylcytosine, more efficiently in ss-DNA than in ds-DNA. Here, we tested the repair of weaker AlkB substrates 1-methylguanine and 3-methylthymine, and found that AlkB prefers to repair them in ds-DNA. We also discovered AlkB and its human homologs, ABH2 and ABH3, are able to repair the aforementioned adducts when the adduct is present in a mismatched base pair. These observations demonstrate the strong adaptability of AlkB on repairing various adducts in different environments.
The DNA and RNA repair protein AlkB removes alkyl groups
from nucleic
acids by a unique iron- and α-ketoglutarate-dependent oxidation
strategy. When alkylated adenines are used as AlkB targets, earlier
work suggests that the initial target of oxidation can be the alkyl
carbon adjacent to N1. Such may be the case with ethano-adenine (EA),
a DNA adduct formed by an important anticancer drug, BCNU, whereby
an initial oxidation would occur at the carbon adjacent to N1. In
a previous study, several intermediates were observed suggesting a
pathway involving adduct restructuring to a form that would not hinder
replication, which would match biological data showing that AlkB almost
completely reverses EA toxicity in vivo. The present study uses more
sensitive spectroscopic methodology to reveal the complete conversion
of EA to adenine; the nature of observed additional putative intermediates
indicates that AlkB conducts a second oxidation event in order to
release the two-carbon unit completely. The second oxidation event
occurs at the exocyclic carbon adjacent to the N6 atom of adenine. The observation of oxidation of a carbon
at N6 in EA prompted us to evaluate N6-methyladenine (m6A), an important epigenetic
signal for DNA replication and many other cellular processes, as an
AlkB substrate in DNA. Here we show that m6A is indeed a substrate
for AlkB and that it is converted to adenine via its 6-hydroxymethyl
derivative. The observation that AlkB can demethylate m6A in vitro
suggests a role for AlkB in regulation of important cellular functions
in vivo.
The HSF (heat shock factor) gene family contains highly conserved plant-specific transcription factors that play an important role in plant high-temperature stress responses. The present study aimed to characterize the HSF transcription factor genes in tomato (Solanum lycopersicum), which is an important vegetable crop worldwide and the model plant for fruit development studies. Twenty-six SlyHSF genes were identified in tomato, and the phylogenetic analysis showed the possible evolution profile of subgroups among in the plant kingdom. A new group O was identified that involved HSF genes in primitive plant species, like in the green algae, mosses and lycophytes. The gene structure and motifs of each SlyHSF were comprehensively analyzed. We identified orthologous, co-orthologous and paralogous HSF gene pairs in tomato, Arabidopsis and rice, and constructed a complex interaction network among these genes. The SlyHSF genes were expressed differentially in different species and at a higher level in mature fruits. The qPCR analysis was performed and showed SlyHSF genes greatly participate in plant heat tolerant pathways. Our comprehensive genome-wide analysis provided insights into the HSF gene family of tomatoes.
Our recent work showed that close relationships result in shared cognitive and neural representations of the self and one's mother in collectivistic individuals (Zhu et al., 2007, Neuroimage, 34, 1310-7). However, it remains unknown whether close others, such as mother, father and best friend, are differentially represented in collectivistic brains. Here, using functional magnetic resonance imaging and a trait judgment task, we showed evidence that, while trait judgments of the self and mother generated comparable activity in the medial prefrontal cortex (MPFC) and anterior cingulate (ACC) of Chinese adults, trait judgments of mother induced greater MPFC/ACC activity than trait judgments of father and best friend. Our results suggest that, while neural representations of the self and mother overlapped in the MPFC/ACC, close others such as mother, father and best friend are unequally represented in the MPFC/ACC of collectivistic brains.
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