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Mechanisms Controlling Germ Plasm Enrichment during Asymmetric Cell Division in the C. elegans Embryo
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| 자료유형 | 학위논문 |
|---|---|
| 서명/저자사항 | Mechanisms Controlling Germ Plasm Enrichment during Asymmetric Cell Division in the C. elegans Embryo. |
| 개인저자 | Gauvin, Timothy Joseph. |
| 단체저자명 | Dartmouth College. Biology. |
| 발행사항 | [S.l.]: Dartmouth College., 2019. |
| 발행사항 | Ann Arbor: ProQuest Dissertations & Theses, 2019. |
| 형태사항 | 132 p. |
| 기본자료 저록 | Dissertations Abstracts International 81-04B. Dissertation Abstract International |
| ISBN | 9781085686310 |
| 학위논문주기 | Thesis (Ph.D.)--Dartmouth College, 2019. |
| 일반주기 |
Source: Dissertations Abstracts International, Volume: 81-04, Section: B.
Advisor: Griffin, Erik E. |
| 이용제한사항 | This item must not be sold to any third party vendors.This item must not be added to any third party search indexes. |
| 요약 | An important task for any embryo during development is to specify the germ cell for ensuring the survival of the species. The nematode, C. elegans, specifies the embryonic germline lineage through the asymmetric enrichment of germ plasm proteins. PIE-1 and POS-1 are two RNA binding proteins that localize to the germ plasm by two well-studied mechanisms: segregation and degradation. It is unknown if other mechanisms contribute to germline fate. Regulation of PIE-1 and POS-1 mobility are required for enrichment to the posterior in the zygote. Two factors that affect mobility are RNA binding and PLK-1 phosphorylation. A previous study has shown that PLK-1 phosphorylation increases POS-1 mobility in the anterior while RNA binding may decrease mobility in the posterior. Currently, there is no evidence to support the direct effects of phosphorylation on RNA binding.In the first part of my dissertation, I show by quantitative microscopy and photobleaching experiments that PIE-1 translation occurs specifically in the germline. To identify potential regulators, I screened ~249 genes by monitoring GFP levels at the 4 cell stage for a change in PIE-1 concentration. I identified two genes, Y-14 and MAG-1 that are part of the exon junction complex (EJC) as novel regulators of maternal translation. Neither gene exhibited zygotic translation. Future studies will identify potentials genes that regulate PIE-1 translation.PLK-1 phosphorylates POS-1 and consequently affects the mobility of POS-1 in the zygote. Based on this model, I developed protocols to purify POS-1 and its phosphorylation mutants using RNA binding assays to test oligomerization. By using fluorescence polarization (FP), the POS-1 zinc finger domain binds with the same affinity as POS-1 N-terminal truncation. However, we are not able to make any conclusions since POS-1 aggregates in a size exclusion column. Thus, we would need to find optimal conditions to prevent aggregation of POS-1 and then test if PLK-1 phosphorylation affects POS-1 RNA binding. |
| 일반주제명 | Biology. Developmental biology. |
| 언어 | 영어 |
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