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Tejas Functions in piRNA Biogenesis Via Nuage Assembly in Drosophila

林, 宇軒 大阪大学

2022.03.24

概要

In Drosophila germline cells, the piRNA amplification, called the ping-pong cycle, occurs at perinuclear structures named ‘nuage’. Two PIWI-clade Argonaute proteins, Aubergine (Aub) and Argonaute3 (Ago3), alternately bind and slice the piRNA precursors and TE transcripts into piRNAs assisted by other components localized to nuage, such as Tejas (Tej) and Spindle- E (Spn-E), which are indispensable for piRNA ping-pong amplification, but the molecular basis of how they cooperate in this processing pathway remains elusive.

In the absence of Tej, high-resolution imaging of ovaries containing endogenously fluorescent-tagged SpnE, Vas, Ago3, and Aub revealed an apparent defect in the localization of several piRNA factors. Core helicase Vas lost its granulation, and the function unknown helicase SpnE displayed a unique nuclear accumulation in tej mutant ovaries. piRNA precursors processing and the proper PIWI proteins piRNA loading is fractured.

I suggested that Tej regulates the dynamics of Spn-E and Vas as an organizer for nuage and potentially sustains the limited contact between the Vas and Spn-E. I newly identified a functional NLS in Spn-E and an Spn-E recruiting motif in Tej that cooperatively controlled the subcellular localization of Spn-E. Further research revealed that distinct parts of Tej engaged in the proper formation of the nuage granules by recruiting different nuage components, which ensure the processing of the piRNA precursors. Tej also regulations the molecular kinetic of the nuage components in vitro by utilizing its disordered region, indicating the contribution of Tej in the dynamic of nuage granules. Taken together, Tej maintains the female Drosophila piRNA biogenesis machinery in the germline cells as a multifunctional organizer of nuage, which takes a vital role in the piRNA precursor processing of germline cells.

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