Abstract

Cap binding protein 80 (Cbp80) is the larger subunit of the nuclear cap-binding complex (nCBC), which is known to play important roles in nuclear mRNA processing, export, stability and quality control events. Reducing Cbp80 mRNA levels in the female germline revealed that Cbp80 is also involved in defending the germline against transposable elements. Combining such knockdown experiments with large scale sequencing of small RNAs further showed that Cbp80 is involved in the initial biogenesis of piRNAs as well as in the secondary biogenesis pathway, the ping-pong amplification cycle. We further found that Cbp80 knockdown not only led to the upregulation of transposons, but also to delocalization of Piwi, Aub and Ago3, key factors in the piRNA biosynthesis pathway. Furthermore, compared to controls, levels of Piwi and Aub were also reduced upon knock down of Cbp80. On the other hand, with the same treatment we could not detect significant changes in levels or subcellular distribution (nuage localization) of piRNA precursor transcripts. This shows that Cbp80 plays an important role in the production and localization of the protein components of the piRNA pathway and it seems to be less important for the production and export of the piRNA precursor transcripts.

Highlights

  • PiRNAs act in a gene silencing mechanism that controls the expression and translocation of transposable genetic elements (TE)

  • An RNAi screen for genes required for repression of transposable elements (TEs) indicated that Cap binding protein 80 (Cbp80) might be required for the repression of different types of TEs [7]

  • Germline cells seem to have a high need for Cbp80 during the period when nos-Gal4 and pCog-Gal4 are expressed in the germarium, but they seem to be less dependent on Cbp80 for their development and survival during the later phases when matα4-GAL4 is active

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Summary

Introduction

PiRNAs act in a gene silencing mechanism that controls the expression and translocation of transposable genetic elements (TE) This activity is important in the germline where it is needed for maintaining genome integrity [1] [2]. PiRNAs form a group of small RNAs, ranging in size from 23 or 24 to 30–32 nucleotides They induce RNA-mediated gene silencing by complementary pairing with target sequences. In Drosophila these are the P element-induced wimpy testis protein (Piwi), Aubergine (Aub) and Argonaute 3 (Ago). In Drosophila these are the P element-induced wimpy testis protein (Piwi), Aubergine (Aub) and Argonaute 3 (Ago3) These three genes and proteins are essential for the production of the piRNAs [1] [2].

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