GSBS Dissertations and Theses

Approval Date

8-2-2013

Document Type

Doctoral Dissertation

Academic Program

Interdisciplinary Graduate Program

Department

Program in Molecular Medicine

First Thesis Advisor

William Theurkauf, PhD

Keywords

DEAD-box RNA Helicases, DNA Transposable Elements, Drosophila Proteins, Drosophila melanogaster, Germ Cells, Nuclear Envelope, Small Interfering RNA

Subjects

Dissertations, UMMS; DEAD-box RNA Helicases; DNA Transposable Elements; Drosophila Proteins; Drosophila melanogaster; Germ Cells; Nuclear Envelope; RNA, Small Interfering

Abstract

Transposon silencing is required to maintain genome stability. The non-coding piRNAs effectively suppress of transposon activity during germline development. In the Drosophila female germline, long precursors of piRNAs are transcribed from discrete heterochromatic clusters and then processed into primary piRNAs in the perinuclear nuage. However, the detailed mechanism of piRNA biogenesis, specifically how the nuclear and cytoplasmic processes are connected, is not well understood. The nuclear DEAD box protein UAP56 has been previously implicated in protein-coding gene transcript splicing and export. I have identified a novel function of UAP56 in piRNA biogenesis. In Drosophila egg chambers, UAP56 co-localizes with the cluster-associated HP1 variant Rhino. Nuage is a germline-specific perinuclear structure rich in piRNA biogenesis proteins, including Vasa, a DEAD box with an established role in piRNA production. Vasa-containing nuage granules localize directly across the nuclear envelope from cluster foci containing UAP56 and Rhino, and cluster transcripts immunoprecipitate with both Vasa and UAP56. Significantly, a charge-substitution mutation that alters a conserved surface residue in UAP56 disrupts co-localization with Rhino, germline piRNA production, transposon silencing, and perinuclear localization of Vasa. I therefore propose that UAP56 and Vasa function in a piRNA-processing compartment that spans the nuclear envelope.

DOI

10.13028/M2GS3J

Rights and Permissions

Copyright is held by the author, with all rights reserved.

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