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Matzke, M.

Publications and source records attributed to Matzke, M..

2 recordsLinked to original sources

Identification and functional categorization of the most stably and most variably expressed genes and retained introns in Arabidopsis thaliana seedlings

We used a large, uniform, RNA-seq dataset to investigate the stability of gene expression and intron retention in Arabidopsis thaliana seedlings. Functional classification of the most stably and most variably expressed genes was determined by GO enrichment analyses. Many variably expressed genes encoded proteins involved in photosynthesis and chloroplast structure, likely reflecting their sensitivity to changeable light intensities, and in stress responses, which allow plants to cope with environmental challenges. As revealed by GO enrichment analysis, the most stably expressed genes were involved in protein, lipid and vesicle trafficking, suggesting that fluctuations in expression of these genes are suboptimal for normal seedling development. GO analyses of genes containing either usually retained or usually spliced introns revealed no consistent enrichments for any specific functional categories. However, highly retained introns were often located in the first or last position, which may contain 5 and 3 UTRs necessary for transcriptional regulation and mRNA transport and stability. Conversely, usually spliced introns were more frequently located in internal portions of pre-mRNAs, indicating that reliable splicing in coding regions is needed to prevent the formation of premature stop codons. The large RNA-seq dataset we generated can be useful for investigating additional aspects of gene expression and pre-mRNA splicing in Arabidopsis seedlings and sets a precedent for future large scale transcriptome analyses.

plant biology↗

A GFP splicing reporter in a coilin mutant background reveals links between alternative splicing, siRNAs and coilin function in Arabidopsis thaliana

Coilin is a scaffold protein essential for the structure of Cajal bodies, which are nucleolar-associated, nonmembranous organelles that coordinate the assembly of nuclear ribonucleoproteins (RNPs) including spliceosomal snRNPs. To learn more about coilin functions and pathways in plants, we conducted a genetic suppressor screen using a coilin (coi1) mutant in Arabidopsis thaliana and performed an immunoprecipitation-mass spectrometry analysis on coilin protein. The coi1 mutations modify alternative splicing of a GFP reporter gene, resulting in a hyper-GFP phenotype in young coi1 seedlings relative to the intermediate wild-type level. As shown here, this hyper-GFP phenotype is extinguished in older coi1 seedlings by posttranscriptional gene silencing triggered by siRNAs derived from aberrant splice variants of GFP pre-mRNA. In the coi1 suppressor screen, we identified suppressor mutations in WRAP53, a putative coilin-interacting protein; SMU2, a predicted splicing factor; and ZC3HC1, an incompletely characterized zinc finger protein. These suppressor mutations return the hyper-GFP fluorescence of young coi1 seedlings to the intermediate wild-type level. Additionally, zc3hc1 coi1 mutants display more extensive GFP silencing and elevated levels of GFP siRNAs, suggesting the involvement of wild-type ZC3HC1 in siRNA biogenesis or stability. The immunoprecipitation-mass spectrometry analysis reinforced coilins roles in pre-mRNA splicing, nucleolar chromatin structure, and rRNA processing. Coilins participation in these processes, at least some of which incorporate small RNAs, supports the hypothesis that coilin acts as a chaperone for small noncoding RNAs. Our study demonstrates the usefulness of the GFP splicing reporter for investigating alternative splicing, ribosome biogenesis, and siRNA-mediated silencing in the context of coilin function.

plant biology↗