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Profiling of cytosine DNA-methylation in the Arabidopsis CAF-1 mutant fas2

ABSTRACT: Profiling of cytosine DNA-methylation in the Arabidopsis CAF-1 mutant fas2

PROVIDER: PRJNA412659 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6121827_1.fastq.gz	Fastqsanger.gz
	SRR6121827_2.fastq.gz	Fastqsanger.gz
	SRR6121828_1.fastq.gz	Fastqsanger.gz
	SRR6121828_2.fastq.gz	Fastqsanger.gz
	SRR6121829_1.fastq.gz	Fastqsanger.gz

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Profiling of cytosine DNA-methylation in the Arabidopsis CAF-1 mutant fas2

Project description:Chromatin Assembly Factor CAF-1 is a histone chaperone involved in nucleosome formation after replication. Here, cytosine DNA-methylation was profiled by bisulphite DNA sequencing in Arabidopsis wild type and the fas2 CAF-1 mutant. To test for transgenerational changes in DNA methylation, fas2 mutants of generation 1 and 6 were compared.

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Project description:Malaria parasites have evolved a well-adjusted developmental program to progress through the complex life cycle in the human and mosquito host. Here we identify cytosine methylation of tRNAAsp (GTC) as being critical to maintain stable protein synthesis under cellular stress. Using conditional knock out of a member of the DNA methyltransferases family called Pf-DNMT2, RNA bisulfite sequencing demonstrates the selective cytosine methylation of this enzyme of tRNAAsp (GTC) at position C38. Although no growth defect on parasite asexual proliferation was observed, Pf-DNMT2 KO parasites show a striking increase in their sexual commitment and an increased sensitivity to Dihydroartemisinin. In mutant parasites we observe a dramatic decrease of tRNAAsp (GTC) when compared to non-stressed parasites and mass spectrometry analysis shows the selective downregulation of proteins with GAC codon bias. Here we identify an epitranscriptomic mechanism that safeguards protein translation and homeostasis of sexual commitment during cellular stress in malaria parasites

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Transcription profiling of Arabidopsis CAF-1 depletion/gamma-irradiation

Project description:CAF-1 is involved in nucleosome assembly following DNA replication and nucleotide excision repair. In Arabidopsis thaliana, the 3 CAF-1 subunits p150, p60 and p48 are encoded by FAS1, FAS2 and, most likely, MSI1, respectively.fas mutants exhibited increased levels of DNA double-strand breaks, a G2 phase retardation, and elevated levels of mRNAs coding for proteins involved in homologous recombinational repair. The same genes that were transcriptionally up-regulated in the fas mutants were also found to have a higher steady state level of transcription in wild-type Arabidopsis plants exposed to gamma-irradiation. Experiment Overall Design: Total RNA prepared from 4-week-old seedlings (without the roots i.e. true leaves with cotyledon and hypocotyls) using an RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Four-week-old sterile seedlings of wild-type plants (ecotype Nossen) were irradiated with 100 Gy of gamma-rays at a dose rate of 300 Gy/h. Six and 12 hours after irradiation, samples were frozen in liquid nitrogen and immediately ground for total RNA isolation. After confirming its quality, extracted RNA was prepared for microarray analysis. An Arabidopsis 2 Oligo Microarray kit (Agilent Technology, CA, USA) was used in this study. After confirming its quality, extracted RNA was prepared for microarray analysis as described by the manufacturer (Agilent) using the suggested chemicals and reagents.

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