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Characterization of Two Opposing Sma...

Wu, Monica.

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Characterization of Two Opposing Small RNA Pathways Required for Germline Homeostasis in Caenorhabditis elegans.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Characterization of Two Opposing Small RNA Pathways Required for Germline Homeostasis in Caenorhabditis elegans./
作者:	Wu, Monica.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2018,
面頁冊數:	219 p.
附註:	Source: Dissertations Abstracts International, Volume: 80-02, Section: B.
Contained By:	Dissertations Abstracts International80-02B.
標題:	Molecular biology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10792769
ISBN:	9780438190498

Characterization of Two Opposing Small RNA Pathways Required for Germline Homeostasis in Caenorhabditis elegans.
Wu, Monica.

Characterization of Two Opposing Small RNA Pathways Required for Germline Homeostasis in Caenorhabditis elegans. - Ann Arbor : ProQuest Dissertations & Theses, 2018 - 219 p.

Source: Dissertations Abstracts International, Volume: 80-02, Section: B.

Thesis (Ph.D.)--University of Toronto (Canada), 2018.

This item is not available from ProQuest Dissertations & Theses.

Central to all small RNA pathways are the Argonaute (AGO) proteins, which interact with small RNAs to regulate gene expression in a sequence-specific manner. The characterization of AGOs across species has demonstrated that AGOs generally interact with different complements of small RNAs to perform distinct regulatory functions. In this thesis, I combined molecular biology and high-throughput sequencing to further characterize two small RNA pathways in Caenorhabditis elegans. First, I utilized the closely-related nematode species Caenorhabditis briggsae as a comparative genomics tool and determined that the CSR-1 pathway is functional in other nematode species. Next, I characterized two additional components of the CSR-1 pathway, CHP-1 and EMB-4, and implicated them in the loading of CSR-1 small RNAs and export of CSR-1 targets from the nucleus to the cytoplasm, respectively. Finally, I set out to characterize, C04F12.1, an AGO that is most closely related to CSR-1. Remarkably, I found that this AGO interacts with multiple classes of small RNAs, including the essential mir-51 family of microRNAs, which is known as the mir-100 in mammals, as well as subsets of piRNAs and CSR-1 22G-RNAs. mRNA and protein expression analyses demonstrated that C04F12.1 is broadly expressed throughout C. elegans development. Immunolocalization studies showed C04F12.1 expression predominantly in somatic tissues with dynamic nuclear localization in embryos. Genome-wide mRNA-seq data implicated C04F12.1 in silencing the expression of a large portion of germline-expressed genes. Furthermore, at elevated temperatures, C04F12.1 mutant embryos possess increased number of cells that aberrantly express germline markers. Thus, we propose a model in which C04F12.1 and its bound 22G-RNAs functions antagonistically to the CSR-1 pathway in somatic cells to silence germline gene expression and maintain somatic cell identity. Collectively, these studies have shed light on how different small RNA pathways can co-regulate mRNA targets to maintain the proper balance of gene expression.

ISBN: 9780438190498Subjects--Topical Terms:

517296
Molecular biology.
Subjects--Index Terms:

Argonaute

Characterization of Two Opposing Small RNA Pathways Required for Germline Homeostasis in Caenorhabditis elegans.
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520 $a Central to all small RNA pathways are the Argonaute (AGO) proteins, which interact with small RNAs to regulate gene expression in a sequence-specific manner. The characterization of AGOs across species has demonstrated that AGOs generally interact with different complements of small RNAs to perform distinct regulatory functions. In this thesis, I combined molecular biology and high-throughput sequencing to further characterize two small RNA pathways in Caenorhabditis elegans. First, I utilized the closely-related nematode species Caenorhabditis briggsae as a comparative genomics tool and determined that the CSR-1 pathway is functional in other nematode species. Next, I characterized two additional components of the CSR-1 pathway, CHP-1 and EMB-4, and implicated them in the loading of CSR-1 small RNAs and export of CSR-1 targets from the nucleus to the cytoplasm, respectively. Finally, I set out to characterize, C04F12.1, an AGO that is most closely related to CSR-1. Remarkably, I found that this AGO interacts with multiple classes of small RNAs, including the essential mir-51 family of microRNAs, which is known as the mir-100 in mammals, as well as subsets of piRNAs and CSR-1 22G-RNAs. mRNA and protein expression analyses demonstrated that C04F12.1 is broadly expressed throughout C. elegans development. Immunolocalization studies showed C04F12.1 expression predominantly in somatic tissues with dynamic nuclear localization in embryos. Genome-wide mRNA-seq data implicated C04F12.1 in silencing the expression of a large portion of germline-expressed genes. Furthermore, at elevated temperatures, C04F12.1 mutant embryos possess increased number of cells that aberrantly express germline markers. Thus, we propose a model in which C04F12.1 and its bound 22G-RNAs functions antagonistically to the CSR-1 pathway in somatic cells to silence germline gene expression and maintain somatic cell identity. Collectively, these studies have shed light on how different small RNA pathways can co-regulate mRNA targets to maintain the proper balance of gene expression.
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