The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival

The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival

Bibliographic Details
Main Author: Zagore, Leah Louise
Language:English
Published: Case Western Reserve University School of Graduate Studies / OhioLINK 2020
Subjects:
Biochemistry
Molecular Biology
RBP
DAZL
RNA regulation
Germ cell development
RNA networks
Post-transcriptional gene regulation
Spermatogenesis
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=case1575647143675768
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-case15756471436757682021-08-03T07:13:36Z The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival Zagore, Leah Louise Biochemistry Molecular Biology RBP DAZL RNA regulation Germ cell development RNA networks Post-transcriptional gene regulation Spermatogenesis RNA binding proteins have essential roles in post-transcriptional gene regulation, evidenced by the fact that abnormal expression of these proteins is linked to many diseases. The main focus of this dissertation is the germ cell-restricted RNA binding protein DAZL (Deleted in Azoospermia Like), a member of the DAZ family (DAZ, DAZL, and BOULE). These proteins were first shown to be essential for spermatogenesis in humans and mice over two decades ago. Thus, while the biological importance of the DAZ proteins is clear, many critical questions have remained, including the identities of their direct RNA targets, how these RNAs are regulated, and why loss of this regulation results in germ cell defects.Here, we generated high-resolution, transcriptome-wide maps of DAZL-RNA interactions in mouse testes. These maps reveal Dazl binding to thousands of mRNAs, predominantly through sequence-specific interactions near the polyA tail. Using Cre-lox technology in transgenic mice and fluorescence activated cell sorting (FACS), we isolated Dazl knockout germ cells and used RNA-Seq to identify mRNAs sensitive to DAZL ablation. Intersection of the RNA-Seq and DAZL-RNA interaction datasets revealed that DAZL enhances expression of a subset of directly-bound transcripts, namely mRNAs for a network of essential cell cycle regulatory genes and a number of genes critical for spermatogenesis. We also demonstrate that DAZL’s polyA-proximal binding is facilitated by interactions between PABPC1 and the mRNA polyA tail, uncovering a novel mechanism for recruitment of an RNA-binding protein to mRNA. Our observations reveal the critical function of DAZL as a master regulator of a specific mRNA program necessary for germ cell survival in mammals. Given the functional conservation of DAZ family proteins, these findings also provide important insights into the molecular basis for azoospermia in 10-15% of infertile men with Y chromosome deletions affecting the DAZ gene. Overall, the work presented in this dissertation critically advances our understanding of the molecular mechanisms that control mammalian germ cell maintenance. 2020-01-24 English text Case Western Reserve University School of Graduate Studies / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=case1575647143675768 http://rave.ohiolink.edu/etdc/view?acc_num=case1575647143675768 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Biochemistry
Molecular Biology
RBP
DAZL
RNA regulation
Germ cell development
RNA networks
Post-transcriptional gene regulation
Spermatogenesis
spellingShingle Biochemistry
Molecular Biology
RBP
DAZL
RNA regulation
Germ cell development
RNA networks
Post-transcriptional gene regulation
Spermatogenesis
Zagore, Leah Louise
The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival
author Zagore, Leah Louise
author_facet Zagore, Leah Louise
author_sort Zagore, Leah Louise
title The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival
title_short The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival
title_full The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival
title_fullStr The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival
title_full_unstemmed The Molecular Function of the RNA Binding Protein DAZL in Male Germ Cell Survival
title_sort molecular function of the rna binding protein dazl in male germ cell survival
publisher Case Western Reserve University School of Graduate Studies / OhioLINK
publishDate 2020
url http://rave.ohiolink.edu/etdc/view?acc_num=case1575647143675768
work_keys_str_mv AT zagoreleahlouise themolecularfunctionofthernabindingproteindazlinmalegermcellsurvival
AT zagoreleahlouise molecularfunctionofthernabindingproteindazlinmalegermcellsurvival
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