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Tetrahymena thermophila

ABSTRACT: Semi-conservative transmission of DNA N6-adenine methylation in a unicellular eukaryote

PROVIDER: PRJNA932808 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16944756_subreads.fastq.gz	Fastqsanger.gz
	SRR16944760_subreads.fastq.gz	Fastqsanger.gz
	SRR16944761_subreads.fastq.gz	Fastqsanger.gz
	SRR16944762_subreads.fastq.gz	Fastqsanger.gz
	SRR16944764_subreads.fastq.gz	Fastqsanger.gz

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Genome-wide analysis of chromatin and transcription in the ciliates Oxytricha trifallax and Tetrahymena thermophila

Project description:DNA N6-adenine methylation (6mA) has recently been reported in diverse eukaryotes, spanning unicellular organisms to metazoans. Yet the functional significance of 6mA remains elusive due to its low abundance, difficulty of manipulation within native DNA, and lack of understanding of eukaryotic 6mA writers. Through biochemical fractionation of nuclear extracts, we identify a putative DNA 6mA methyltransferase in ciliates, termed MTA1. It contains an MT-A70 domain but is phylogenetically distinct from all known RNA and DNA methyltransferases. Disruption of MTA1 leads to the genome-wide loss of 6mA in vegetative cells and abolishment of the consensus ApT dimethylated motif in vivo. Genes exhibit subtle changes in chromatin organization or RNA expression upon 6mA loss, depending on their starting methylation level. Mutants fail to complete the sexual cycle, which normally coincides with a peak of MTA1 expression. Thus, MTA1 functions in a developmental stage-specific manner. We then determine the impact of 6mA on chromatin organization in vitro by reconstructing full-length ciliate chromosomes harboring 6mA in native or ectopic positions. Using these synthetic chromosomes, we show that 6mA directly disfavors nucleosomes in a local, quantitative manner in vitro, independent of DNA sequence. Furthermore, the chromatin remodeler ACF can overcome this effect. Our study identifies a novel MT-A70 protein necessary for eukaryotic 6mA methylation and defines the impact of 6mA on chromatin organization using epigenetically defined chromosomes.

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Functional analysis of the methyltransferase SMYD in the single-cell model organism Tetrahymena thermophila

Project description:In order to resolve controversies concerning SMYD's substrates and functions, we studied SMYD1 (TTHERM_00578660), the only homologue in the unicellular eukaryote Tetrahymena thermophila. We epitope-tagged SMYD1, and analyzed its localization and interactome. We also characterized ΔSMYD1 cells, focusing on the replication and transcription phenotype. Our results show that: 1) SMYD1 is present in both cytoplasm and transcriptionally active macronucleus and shuttles between cytoplasm and macronucleus, which is consistent with its association with both histone and non-histone substrates; 2) SMYD1 is involved in DNA replication and regulates transcription of metabolism-related genes; 3) HSP90 is an potential substrate for SMYD1 and it may regulate target selection of HSP90, leading to pleiotropic effects in both the cytoplasm and the nucleus.

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RNA-seq analysis of Tetrahymena thermophila

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Tetrahymena thermophila SB210

Project description:Dual modes of DNA N6-methyladenine maintenance by 2 distinct methyltransferase complexes

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