Dataset Information

Grape ASR-Silencing Sways Nuclear Proteome, Histone Marks and Interplay of Intrinsically Disordered Proteins.

ABSTRACT: In order to unravel the functions of ASR (Abscisic acid, Stress, Ripening-induced) proteins in the nucleus, we created a new model of genetically transformed grape embryogenic cells by RNAi-knockdown of grape ASR (VvMSA). Nuclear proteomes of wild-type and VvMSA-RNAi grape cell lines were analyzed by quantitative isobaric tagging (iTRAQ 8-plex). The most significantly up- or down-regulated nuclear proteins were involved in epigenetic regulation, DNA replication/repair, transcription, mRNA splicing/stability/editing, rRNA processing/biogenesis, metabolism, cell division/differentiation and stress responses. The spectacular up-regulation in VvMSA-silenced cells was that of the stress response protein VvLEA D-29 (Late Embryogenesis Abundant). Both VvMSA and VvLEA D-29 genes displayed strong and contrasted responsiveness to auxin depletion, repression of VvMSA and induction of VvLEA D-29. In silico analysis of VvMSA and VvLEA D-29 proteins highlighted their intrinsically disordered nature and possible compensatory relationship. Semi-quantitative evaluation by medium-throughput immunoblotting of eighteen post-translational modifications of histones H3 and H4 in VvMSA-knockdown cells showed significant enrichment/depletion of the histone marks H3K4me1, H3K4me3, H3K9me1, H3K9me2, H3K36me2, H3K36me3 and H4K16ac. We demonstrate that grape ASR repression differentially affects members of complex nucleoprotein structures and may not only act as molecular chaperone/transcription factor, but also participates in plant responses to developmental and environmental cues through epigenetic mechanisms.

SUBMITTER: Atanassov H

PROVIDER: S-EPMC8835812 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature

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Grape ASR-Silencing Sways Nuclear Proteome, Histone Marks and Interplay of Intrinsically Disordered Proteins.

Atanassov Hristo H Parrilla Jonathan J Artault Caroline C Verbeke Jérémy J Schneider Thomas T Grossmann Jonas J Roschitzki Bernd B Atanassova Rossitza R

International journal of molecular sciences 20220128 3

In order to unravel the functions of ASR (Abscisic acid, Stress, Ripening-induced) proteins in the nucleus, we created a new model of genetically transformed grape embryogenic cells by RNAi-knockdown of grape ASR (VvMSA). Nuclear proteomes of wild-type and VvMSA-RNAi grape cell lines were analyzed by quantitative isobaric tagging (iTRAQ 8-plex). The most significantly up- or down-regulated nuclear proteins were involved in epigenetic regulation, DNA replication/repair, trans ...[more]

PMID: 35163458

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Project description:The biological roles of ASRs (Abscisic acid, Stress, Ripening proteins) involved in certain crucial switches of plant development (grain and pollen germination, senescence, fruit ripening), and responses to environmental cues (drought, salinity, cold, nutrient depletion, fungus infection, heavy metal toxicity) remain still elusive. The functional duality of ASRs, as chaperones for direct protection of biological macromolecules, and as transcriptional regulators of gene expression, needs in-depth understanding of the underlying mechanisms. To further elucidate the biological functions of ASRs a quantitative proteomic analysis (iTRAQ) was performed comparing the nuclear proteins of grapevine embryogenic 41B cells: wild-type (WT) and genetically modified VvMSA-RNAi cells, silenced for grape ASR. Among all identified proteins, thirty-one were differentially expressed in WT and VvMSA-silenced cells and may be classified into five functional groups: metabolism, stress responses, ribosome biogenesis, transcription and chromatin architecture/remodeling. The impact of VvMSA silencing was also confirmed at the level of the genes encoding the differentially expressed nuclear proteins, and the revealed most pronounced effects at protein level suggest a more direct correlation to the putative biological function. Remarkably, many of the thirty-one differentially expressed proteins are supposedly involved in epigenetic events. To further corroborate this hypothesis immunoblot assay of Multiscreen type was performed using twenty-one antibodies to different post-translational modifications (PTMs) of histones H3 and H4 in both cell types, WT and VvMSA-RNAi. The subsequent ImageQuant analysis revealed marked semi-quantitative differences in most PTMs of grape histones H3 and H4, with sharp increase of H3K27ac, H3K36me3, H4K5ac and H4K12ac, as well as a strong decrease of H3K27me3 in VvMSA-RNAi cells. These findings provide new insights on a plausible role of VvMSA as an ASR protein involved in epigenetic mechanisms

Dataset Information

Grape ASR-Silencing Sways Nuclear Proteome, Histone Marks and Interplay of Intrinsically Disordered Proteins.

Publications

Grape ASR-Silencing Sways Nuclear Proteome, Histone Marks and Interplay of Intrinsically Disordered Proteins.

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