Vaid, N. | Search bioRxiv

bioRxiv2017

AtRsgA from Arabidopsis thaliana controls maturation of the small subunit of the chloroplast ribosome

SummaryPlastid ribosomes are very similar in structure and function to ribosomes of their bacterial ancestors. Since ribosome biogenesis is not thermodynamically favourable at biological conditions, it requires activity of many assembly factors. Here, we have characterized a homolog of bacterial rsgA in Arabidopsis thaliana and show that it can complement the bacterial homolog. Functional characterization of a strong mutant in Arabidopsis revealed that the protein is essential for plant viability, while a weak mutant produced dwarf, chlorotic plants that incorporated immature pre-16S ribosomal RNA into translating ribosomes. Physiological analysis of the mutant plants revealed smaller, but more numerous chloroplasts in the mesophyll cells, reduction of chlorophyll a and b, depletion of proplastids from the rib meristem and decreased photosynthetic electron transport rate and efficiency. Comparative RNA-sequencing and proteomic analysis of the weak mutant and wild-type plants revealed that various biotic stress-related, transcriptional regulation and post-transcriptional modification pathways were repressed in the mutant. Intriguingly, while nuclear- and chloroplast-encoded photosynthesis-related proteins were less abundant in the mutant, the corresponding transcripts were upregulated, suggesting an elaborate compensatory mechanism, potentially via differentially active retrograde signalling pathways. To conclude, this study reveals a new chloroplast ribosome assembly factor and outlines the transcriptomic and proteomic responses of the compensatory mechanism activated during decreased chloroplast function.\n\nSignificance statementAtRsgA is an assembly factor necessary for maturation of the small subunit of the chloroplast ribosome. Depletion of AtRsgA leads to dwarfed, chlorotic plants and smaller, but more numerous chloroplasts. Large-scale transcriptomic and proteomic analysis revealed that chloroplast-encoded and - targeted proteins were less abundant, while the corresponding transcripts were upregulated in the mutant. We analyse the transcriptional responses of several retrograde signalling pathways to suggest a mechanism underlying this compensatory response.

molecular biology↗

bioRxiv2017

EnsembleNet: ensemble gene function predictions for Arabidopsis thaliana

Despite increasing availability of sequenced genomes, accurate characterization of gene functions is needed to close the genotype-phenotype gap. Recent advances in gene function prediction rely on ensemble approaches that integrate the results from multiple inference methods to produce superior predictions. Yet, these developments remain largely unexplored in plants. We present Neighbor Counting Ensemble, a gene function prediction method which integrates eleven gene co-function networks for Arabidopsis thaliana, and produces more accurate gene function predictions for a larger fraction of genes with unknown function. We used these predictions to identify genes involved in mitochondrial complex I formation, and for five of them we confirmed the predictions experimentally. The ensemble predictions are provided as a user-friendly online database, EnsembleNet, available at http://www.gene2function.de/ensemblenet.html.

bioinformatics↗