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The chromatin-regulating CoREST complex is animal specific and essential for development in the cnidarian Nematostella vectensis

ABSTRACT: Using the sea anemone Nematostella vectensis as a model for early branching metazoans, we show that CoREST is an animal-specific protein that assembles a conserved histone-modifying complex including Lsd1 and HDAC1/2. We further show that the Nematostella complex is similar in composition to vertebrates.

INSTRUMENT(S):

ORGANISM(S): Nematostella Vectensis

TISSUE(S): Embryo

SUBMITTER: Maria Hernandez-Valladares

LAB HEAD: Fabian Rentzsch

PROVIDER: PXD033068 | Pride | 2022-08-08

REPOSITORIES: Pride

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			Action	DRS
	Control_IP.raw	Raw
	Control_IP_05.raw	Raw
	Eclipse_270821_JamesGahan_Control_1.raw	Raw
	Eclipse_270821_JamesGahan_Control_2.raw	Raw
	Eclipse_270821_JamesGahan_Control_3.raw	Raw

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Project description:Transcriptome analysis was used to investigate the global stress response of the Gram-positive bacterium Bacillus subtilis caused by overproduction of the well-secreted AmyQ α-amylase from Bacillus amyloliquefaciens. Analyses of the control and overproducing strains were carried out at the end of exponential growth and in stationary phase, when protein secretion from B. subtilis is optimal. Among the genes that showed increased expression were htrA and htrB, which are part of the CssRS regulon that responds to high-level protein secretion and heat stress. The analysis of the transcriptome profiles of a cssS mutant compared to the wild-type, under identical secretion stress conditions, revealed several genes with altered transcription in a CssRS-dependent manner, for example citM, ylxF, yloA, ykoJ and several genes of the flgB operon. However, a high affinity CssR-binding was only observed for htrA and htrB, and possibly for citM. In addition, the DNA macroarray approach reveal that several genes of the sporulation pathway are downregulated by AmyQ overexpression, and a group of motility-specific (σD-dependent) transcripts were clearly upregulated. Subsequent flow cytometric analyses demonstrate that upon overproduction of AmyQ as well as a non-secretable variant of the α-amylase, the process of sporulation is severely inhibited. The same experiments were implemented to investigate the expression levels of the hag promoter, a well-established reporter for σD-dependent gene expression. This approach confirmed the observations based on our DNA macroarray analyses and led us to conclude that expression levels of several genes involved in motility are maintained at high levels under all conditions of α-amylase overproduction. Secretion stress was applied by overproducing the well-secreted AmyQ α-amylase (pKTH10 vector) from B. amyloliquefaciens. Besides examining secretion stress in wild-type cells, we compared transcriptome profiles of a cssS mutant strain under conditions of high-level AmyQ production. Samples for transcriptome analyses were collected at the late exponential growth stage (one hour before the transition point) and 3 hours upon entry in the stationary growth phase. Three independent cultures of each strain were used and cells were sampled for macroarray experiments. Duplicate spots were averaged in Array-Pro software (Media Cybernetics, Inc.) and the signal was normalized after background subtraction by calculation of the percentage of total signal per gene using Microsoft Excel.

Dataset Information

The chromatin-regulating CoREST complex is animal specific and essential for development in the cnidarian Nematostella vectensis

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