Dataset Information

For euchromatic repeats RID is required for both Repeat Induced Point mutation and nucleation of a novel transitional heterochromatic state

ABSTRACT: Gene inactivation via heterochromatization is important for genome integrity and prevention of unwanted transcripts with dysregulation in cancer. Repeat sequences are subject to heterochromatin inactivation and, in Neurospora, Repeat-Induced-Point mutation (RIP). The initiating factors behind both are poorly understood. We resolve the paradoxical observation that newly introduced Repeat-Linker-Repeat (RLR) constructs require RID alone for RIP, while genomic repeats are RIPed in the absence of RID, showing that eu- and hetero- chromatic repeats are handled differently, the latter requiring DIM-2 as well. The differences between mechanisms associated with older and newer duplicates caution against extrapolation from mechanisms inferred from model experimental systems to all genomically relevant processes. Additionally, while chromatin status affects RIP, RID we also show via lexA tethering is the nucleation centre for a transition from eu- to hetero- chromatin, in an HDA-1 dependent fashion. Constitutive heterochromatin by contrast is HDA1 independent and depends on HDA-1 paralogs. RID is thus a dual function initiator of both RIP and the transition to heterochromatin.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Neurospora Crassa Or74a

TISSUE(S): Ascocarp

SUBMITTER: Ju Huang

LAB HEAD: Ju Huang

PROVIDER: PXD060025 | Pride | 2025-03-28

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	RID_as1_1_PeptideSummary.txt	Txt
	RID_as1_1_ProteinSummary.txt	Txt
	RID_as1_1__FDR.xlsx	Xlsx
	RID_as1_PeptideSummary.txt	Txt
	RID_as1_ProteinSummary.txt	Txt

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Project description:Aspergillus flavus first gained scientific attention for its production of aflatoxin, the most potent naturally occurring toxin and hepatocarcinogenic secondary metabolite. For several decades, The DNA methylation status of A. flavus remains to be controversial. We first applied bisulfite sequencing, the gold standard at present, in conjunction with a biological replicate strategy to investigate the DNA methylation profiling of A. flavus genome. Our results reveal that the DNA methylation level of this fungus turns out to be negligible, comparable to the unmethylated lambda DNA we set as the false positive control of our bisulfite experiments. When comparing the DNA methyltransferase homolog of A. flauvs with that from several selected hypermethylated speices, we find that the DNA methyltransferase homolog of A.flavus as well as the other Aspergillus members groups closely with the RID from Neurospora crassa and Masc1 from Ascobolus immerses, which has been reported as DMT-incapable, but it diverges distantly from the other capable DNA methyltransferases. We observe significant depletion of repeat components within the A. flavus, which may possibly explain the lack of DNA methylation in this fungus. What's more, the RIP-index of the repeat of A. flavus turns out to be higher than the fungi without RID-like enzyme, suggesting this asexual fungus may possibly possess RIP process during the obscure sexual-stage which is very evanescent and may potentially related to DNA methylation. This work contributes to our understanding on the DNA methylation status of A. flavus. Also, it reinforces our views on the DNA methylation in fungal species. What's more, our strategy of applying bisulfite sequencing to DNA methylation detection on species with low DNA methylation may serve as a reference for later scientific investigations on other hypomethylated species. Two replicates were subjected to bisulfite conversion independently, unmethylated lambda DNA as a false positive control is added to both replicates.

Dataset Information

For euchromatic repeats RID is required for both Repeat Induced Point mutation and nucleation of a novel transitional heterochromatic state

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