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Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (BS-RNA-Seq)

ABSTRACT: Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (BS-RNA-Seq)

PROVIDER: PRJNA1186075 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR31348090.fastq.gz	Fastqsanger.gz
	SRR31348091.fastq.gz	Fastqsanger.gz
	SRR31348092.fastq.gz	Fastqsanger.gz
	SRR31348093.fastq.gz	Fastqsanger.gz
	SRR31348094.fastq.gz	Fastqsanger.gz

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Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (RNA-Seq)

Project description:Spermatozoid’s flagella assemble in transcriptionally silent spermatids and thus depends on post-transcriptional regulation of gene expression. Mutations in Nsun7 gene are known to cause male infertility in human and mice. We identified m5C-specific Nsun7 RNA methyltransferase as a protein present in elongated spermatids and interacting with RNAs specific for this type of spermatozoid’s precursor cells. Inactivation of Nsun7 gene in mice leads to upregulation of its RNA interactors, thus indicating that Nsun7 downregulates a set of RNAs in the elongated spermatids. A physiologic consequence of Nsun7 gene knockout is male infertility, which is mechanistically explained by the observed mispositioning of longitudinal columns relative to the axonemal microtubular doublets leading to motility defect.

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Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (CLIP-Seq)

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Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (BS-RNA-Seq)

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Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (CLIP-Seq)

Project description:Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (CLIP-Seq)

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Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (RNA-Seq)

Project description:Positioning of sperm tail longitudinal columns depend on Nsun7, an RNA binding protein destabilizing elongated spermatid transcripts (RNA-Seq)

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NSUN7 is an RNA:m5C catalytically inactive protein that regulates sperm flagellum assembly

Project description:Defects in Nsun7, a member of the main RNA:m5C modifying enzymes, known as the NOL1/NOP2/Sun domain (NSUN) family, cause male infertility. The NSUN family includes NSUN1-7, all possessing conserved motifs IV and VI essential for RNA:m5C formation. However, due to the controversy regarding the catalytic activity and RNA substrates of NSUN7, the pathogenic mechanism underlyingof NSUN7-related male infertility remains unclear. Here, we showed that NSUN7 is predominantly enriched in adult mouse testes, particularly in elongated spermatids. Through m5C mass spectrometry detection and bisulfite sequencing, we found that knockout of Nsun7 has no impact on testicular RNA:m5C modification. Mechanistically, NSUN7 loses its ability to bind S-adenosylmethionine (SAM) due to sequence variations in motif IV compared towith the conserved residues observed in NSUN1-6. Notably, the key SAM-binding Asp in motif IV is replaced by Leu in NSUN7, thereby abolishing SAM binding. Nsun7 deficiency impairs sperm progressive motility, accompanied by defects in axonemes and mispositioning of longitudinal columns. Single-cell RNA sequencing showed that knockout of Nsun7 results in a significantly decrease in the levels of a cohort of mRNAs related to cilium organization in elongated spermatids. Collectively, our study demonstrateds that NSUN7 is the only RNA:m5C catalytically inactive member in the NSUN family, and that it regulates sperm flagellum assembly by modulating the expression of cilium organization-associated genes.

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Proteome of wild type and Nsun7 knockout mice testis

Project description:Investigation of a new RNA-modification-based mechanism in mammalian spermatogenesis. We created a mice line devoid of testis-specific m5C RNA methyltransferase Nsun7. These mice demonstrated male infertility due to a peculiar defect in sperm tail assembly – mispositioning of the longitudinal columns of the fibrous sheath. The whole testis proteomes of wild type and Nsun7 knockout mice were obtained to study the molecular pathways affected by NSUN7.

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Small non-coding RNA expression in mouse elongated spermatids

Project description:In order to discover novel small RNAs expressed in elongated spermatids, we isolated elongated spermatids from mouse testis. The small RNA fraction (18-40nt) was cloned and sequenced from total RNA. RNAs extracted from elongated spermatids were used for high throughput sequencing analysis

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