Plants respond and adapt to drought, cold and high-salinity stresses in order to survive. In this study, we applied Arabidopsis Affymetrix tiling arrays to study the whole genome transcriptome under drought, cold, high-salinity and ABA treatment conditions. The bioinformatic analysis using the tiling array data showed that 7,719 non-AGI transcriptional units (TUs) exist in the unannotated "intergenic" regions of Arabidopsis genome. These include 1,275 and 181 TUs that are induced and downregulated, respectively, by the stress or ABA treatments. Most of the non-AGI TUs are hypothetical non-protein-coding RNAs. About 80% of the non-AGI TUs belong to pairs of the fully overlapping sense-antisense transcripts (fSATs). Significant linear correlation between the expression ratios (treated/untreated) of the sense TUs and the ratios of the antisense TUs was observed in the SATs of AGI code/non-AGI TU. We studied the biogenesis mechanisms of the stress- or ABA-inducible antisense RNAs and found that the expression of sense TUs is necessary for the stress- or ABA-inducible expression of the antisense TUs in the fSATs (AGI code/non-AGI TU).

Genome integrity is continuously threatened by external stresses and endogenous hazards such as DNA replication errors and reactive oxygen species. The DNA damage checkpoint in metazoans ensures genome integrity by delaying cell-cycle progression to repair damaged DNA or by inducing apoptosis. ATM and ATR (ataxia-telangiectasia-mutated and -Rad3-related) are sensor kinases that relay the damage signal to transducer kinases Chk1 and Chk2 and to downstream cell-cycle regulators. Plants also possess ATM and ATR orthologs but lack obvious counterparts of downstream regulators. Instead, the plant-specific transcription factor SOG1 (suppressor of gamma response 1) plays a central role in the transmission of signals from both ATM and ATR kinases. Here we show that in Arabidopsis, endoreduplication is induced by DNA double-strand breaks (DSBs), but not directly by DNA replication stress. When root or sepal cells, or undifferentiated suspension cells, were treated with DSB inducers, they displayed increased cell size and DNA ploidy. We found that the ATM-SOG1 and ATR-SOG1 pathways both transmit DSB-derived signals and that either one suffices for endocycle induction. These signaling pathways govern the expression of distinct sets of cell-cycle regulators, such as cyclin-dependent kinases and their suppressors. Our results demonstrate that Arabidopsis undergoes a programmed endoreduplicative response to DSBs, suggesting that plants have evolved a distinct strategy to sustain growth under genotoxic stress.root meristem | protein degradation D amaged DNA needs to be repaired to prevent loss or incorrect transmission of genetic information. Eukaryotic DNA damage checkpoints delay or arrest the cell cycle to provide time for DNA repair before the cell enters a new round of DNA replication or mitosis (1). In metazoans, ATM and ATR (ataxia-telangiectasia-mutated and -Rad3-related) are sensor kinases that play a crucial role in the checkpoint system. ATM specifically responds to DNA double-strand breaks (DSBs), and ATR primarily senses replication stress caused by a persistent block of replication fork progression. ATM deficiency confers hypersensitivity to ionizing radiation (2), whereas ATR knockout mutation is lethal (3, 4), and dominant-negative cell lines display hypersensitivity to UVB light, gamma radiation, hydroxyurea (HU), and aphidicolin (5, 6). ATM and ATR relay the damage signal to transducer kinases Chk2 and Chk1, respectively, which then amplify the signal and regulate an overlapping set of substrates that trigger cell-cycle arrest and DNA repair (1). The transcription factor p53, cyclin-dependent kinase (CDK) inhibitor p21, and Cdc25 phosphatase are downstream regulators that control cell-cycle arrest in response to DNA damage.Comparative sequence analyses among plants, yeast, and animals indicate that some of the factors involved in DNA damage checkpoint and DSB repair systems are conserved between vertebrates and plants (7). Plants also possess ATM and ATR orthologs, and knockout mutants show similar phenotypes...

The nonsense-mediated mRNA decay (NMD) pathway is a wellknown eukaryotic surveillance mechanism that eliminates aberrant mRNAs that contain a premature termination codon (PTC). The UP-Frameshift (UPF) proteins, UPF1, UPF2, and UPF3, are essential for normal NMD function. Several NMD substrates have been identified, but detailed information on NMD substrates is lacking. Here, we noticed that, in Arabidopsis, most of the mRNA-like nonprotein-coding RNAs (ncRNAs) have the features of an NMD substrate. We examined the expression profiles of 2 Arabidopsis mutants, upf1-1 and upf3-1, using a whole-genome tiling array. The results showed that expression of not only protein-coding transcripts but also many mRNA-like ncRNAs (mlncRNAs), including natural antisense transcript RNAs (nat-RNAs) transcribed from the opposite strands of the coding strands, were up-regulated in both mutants. The percentage of the up-regulated mlncRNAs to all expressed mlncRNAs was much higher than that of the upregulated protein-coding transcripts to all expressed proteincoding transcripts. This finding demonstrates that one of the most important roles of NMD is the genome-wide suppression of the aberrant mlncRNAs including nat-RNAs.ncRNA ͉ tiling array ͉ UPF1 ͉ UPF3 In Arabidopsis thaliana, hundreds of nonprotein-coding RNA (ncRNA) transcripts have been discovered based on the cloning of full-length cDNAs, whole-genome tiling array, and deep-sequencing analysis (1-8). They include microRNA (miRNA) precursors, transacting siRNA (ta-siRNA) precursors, and mRNA-like ncRNAs (mlncRNA). The mlncRNAs were divided into 2 types-natural antisense transcript RNAs (natRNAs) that arise from the strands opposite the coding strands and other mlncRNAs.Nonsense-mediated mRNA decay (NMD) is a eukaryotic mRNA quality-control mechanism that eliminates aberrant mRNAs containing a premature termination codon (PTC) from cells to avoid the production of truncated proteins (9-13). Such transcripts can arise by genomic frameshifts, nonsense mutations, inefficiently spliced premRNAs, and so on. Several proteins involved in NMD have already been discovered (10). Among them, the UP-Frameshift (UPF) proteins, UPF1, UPF2, and UPF3, are core components of mRNA surveillance complexes and are essential for normal NMD function. In yeasts and mammals, aberrant transcripts with nonsense mutations are recognized by the UPF complex and then degraded from the 5Ј and 3Ј ends by recruiting-decapping and 5Ј33Ј exonuclease activities and deadenylating and 3Ј35Ј exonuclease activities (14-16).Plants also have a sophisticated NMD system (10). The Arabidopsis genome encodes homologues of 3 UPF genes (UPF1, UPF2, and UPF3). Some UPF1 and UPF3 mutants were obtained in Arabidopsis and analyzed to investigate the NMD system in plants and to identify several mRNA substrates for NMD by using microarrays (17-20). For example, the aberrant mRNAs containing PTC were overaccumulated in the upf1 and upf3 mutants (17)(18)(19).In plants, aberrant mRNAs with termination codons located distant (м300 nt) f...

Quality assurance and correct taxonomic affiliation of data submitted to public sequence databases have been an everlasting problem. The DDBJ Fast Annotation and Submission Tool (DFAST) is a newly developed genome annotation pipeline with quality and taxonomy assessment tools. To enable annotation of ready-to-submit quality, we also constructed curated reference protein databases tailored for lactic acid bacteria. DFAST was developed so that all the procedures required for DDBJ submission could be done seamlessly online. The online workspace would be especially useful for users not familiar with bioinformatics skills. In addition, we have developed a genome repository, DFAST Archive of Genome Annotation (DAGA), which currently includes 1,421 genomes covering 179 species and 18 subspecies of two genera, Lactobacillus and Pediococcus, obtained from both DDBJ/ENA/GenBank and Sequence Read Archive (SRA). All the genomes deposited in DAGA were annotated consistently and assessed using DFAST. To assess the taxonomic position based on genomic sequence information, we used the average nucleotide identity (ANI), which showed high discriminative power to determine whether two given genomes belong to the same species. We corrected mislabeled or misidentified genomes in the public database and deposited the curated information in DAGA. The repository will improve the accessibility and reusability of genome resources for lactic acid bacteria. By exploiting the data deposited in DAGA, we found intraspecific subgroups in Lactobacillus gasseri and Lactobacillus jensenii, whose variation between subgroups is larger than the well-accepted ANI threshold of 95% to differentiate species. DFAST and DAGA are freely accessible at https://dfast.nig.ac.jp.