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Transcriptome analysis of isoxaben-induced loss of cell wall integrity in Arabidopsis seedling liquid culture

ABSTRACT: Plant cells are surrounded by walls, which must often meet opposing functional requirements during plant growth and defense. The cells meet them by modifying wall structure and composition in a tightly controlled and adaptive manner. The modifications seem to be mediated by a dedicated cell wall integrity (CWI) maintenance mechanism. Currently the mode of action of the mechanism is not understood and it is unclear how its activity is coordinated with established plant defense signaling. We investigated both the responses to cell wall damage (CWD) compromising CWI and the underlying mechanism in Arabidopsis thaliana. A cellulose biosynthesis inhibitor isoxaben was used as a tool to induce the loss of cell wall integrity. Isoxaben was chosen because it only affects a certain cell type / differentiation stage, and weakens the cell walls indirectly by inhibiting a biosynthetic process, making CWD occurrence dependent on high turgor levels (allowing simultaneous manipulation of the responses by addition of osmotica like sorbitol, mannitol, etc.). Isoxaben treatment causes structural damage, induction of lesion formation, cell death, deposition of lignin and callose as well as production of jasmonic acid and salicylic acid. Isoxaben-resistant mutant ixr1-1 is included to ensure the specificity of the treatment.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE108987 | GEO | 2018/01/11

SECONDARY ACCESSION(S): PRJNA429347

REPOSITORIES: GEO

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Project description:The Neurospora crassa GUL-1 is part of the COT-1 pathway, which plays key roles in regulating polar hyphal growth and cell wall remodeling. We show that GUL-1 is a bona fide mRNA binding protein (RBP) that can associate with 828 “core” mRNA species. When cell wall integrity (CWI) is challenged, the repertoire of associated RNA species increases up to 2628 mRNAs (including its own transcript). GUL-1 can bind mRNAs of genes related to translation, cell wall remodeling, circadian clock, endoplasmatic reticulum (ER), as well as CWI and osmoregulatory response MAPK pathway components. GUL-1 interacts with over 100 different proteins, including stress granule proteins, components of the translational and cell wall remodeling machinery, as well as ER components and those of the MAPK, COT-1 and STRIPAK complexes. Several additional RBPs were also shown to physically interact with GUL-1. We demonstrate that under stress conditions GUL-1 can localize to the ER and that GUL-1 affects the CWI pathway as is evident via altered phosphorylation levels of MAK-1, interaction with mak-1 transcript and involvement in the expression level of the transcription factor adv-1. Taken together, we suggest that the RBP GUL-1 functions in multiple cellular processes, including the regulation of cell wall remodeling. The latter is mechanistically concerted with the MAK-1 pathway in a stress-related manner. Methods: In this study we show that GUL-1 is a bona fide mRNA binding protein (RBP) that associates with different mRNAs, including its own transcript, as determined by RNA antisense purification and RNA immunoprecipitation experiments. Results: We demonstrate that GUL-1 may physically interact with multiple proteins included translation processes, cell wall remodeling and cell wall integrity (CWI). We show that GUL-1 affects the CWI pathway as is evident via altered phosphorylation levels of MAK-1, interaction with mak-1 transcript and involvement in the expression level of the transcription factor adv-1. Conclusions:We suggest that the RBP GUL-1 functions in the regulation of cell wall remodeling in concert with the MAK-1 pathway in a stress-related manner.

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Transcriptome analysis of isoxaben-induced loss of cell wall integrity in Arabidopsis seedling liquid culture

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