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RNA associating with PIWI proteins in Bombyx mori BmN4 cells

ABSTRACT: RNA associating with PIWI proteins in Bombyx mori BmN4 cells

PROVIDER: PRJNA632342 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11781557.fastq.gz	Fastqsanger.gz
	SRR11781558.fastq.gz	Fastqsanger.gz
	SRR11781559.fastq.gz	Fastqsanger.gz
	SRR11781560.fastq.gz	Fastqsanger.gz
	SRR11781561.fastq.gz	Fastqsanger.gz

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RNA associating with PIWI proteins in Bombyx mori BmN4 cells

Project description:RNAs associating with PIWI proteins were Immunoisolated from BmN4 cells. Sequence libraries were generated with NEBNext Small RNA Library Prep Set for Illumina(NEB). Libraries were sequenced using Illumina MiSeq (single-end, 51 cycles).

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Project description:In germ cells, piRNAs are amplified through the Ping-Pong cycle that depends on reciprocal Slicer-mediated target RNA cleavage by two PIWI members. A germ-specific DEAD-box protein Vasa is required for the process. However, Vasa’s function is poorly understood. Here, we show that target RNAs cleaved by a Bombyx mori (silkworm) PIWI, Siwi, remain to be bound with the protein upon cleavage, but are released in the presence of Vasa in B. mori (BmVasa) and ATP. Under normal conditions, BmVasa co-purifies with Siwi, but not with second B. mori PIWI BmAgo3. However, when BmVasa loses the ATP-binding and RNA-unwinding activities, BmVasa avidly associates with Siwi and BmAgo3, which contains transposon transcripts predominantly in sense orientation, the sources of BmAgo3-piRNAs. Without BmVasa, BmAgo3 is devoid of piRNAs. Thus, BmVasa actively releases target RNAs from Siwi, upon its cleavage, to urge BmAgo3-piRNA complex formation in the Ping-Pong cycle, enabling continuous supply of piRNAs in germ cells.

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Protein-protein interactions of GTSF1L in Bombyx mori

Project description:A mass spectrometry-based proteomics analysis was performed to study the protein binders of GTSF1L in germ cell culture (BmN4; ovary-derived) of Bombyx mori. Anti-HA pull-downs were performed in BmN4 cells transfected with the HA-BmGtsf1L plasmid or the HA-eGFP control plasmid. Proteins were digested in-gel by trypsin. The resultant peptides were then dimethyl labelled and combined. Peptides were measured on a Q Exactive Plus Orbitrap mass spectrometer.

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Genome-wide Identification of Polycomb Target Genes Reveals a Functional Association of Pho with Scm in Bombyx mori

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Project description:Changes in gene expressions associated with egg diapause in multivoltine silkworm Bombyx mori

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