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dc.contributor.authorSarkissian, Madathia
dc.date2022-08-11T08:08:41.000
dc.date.accessioned2022-08-23T16:04:05Z
dc.date.available2022-08-23T16:04:05Z
dc.date.issued2004-07-12
dc.date.submitted2007-04-18
dc.identifier.urihttp://hdl.handle.net/20.500.14038/31620
dc.descriptionIn the process of seeking author's permission to provide full text.
dc.description.abstractFully grown oocytes' of the African clawed frog, Xenopus laevis, are arrested at the diplotene stage of meiotic prophase I, which resembles the G2 phase of the mitotic cell cycle. Re-entry into the meiotic divisions is initiated by hormonal signaling normally provided by progesterone. Progesterone signaling leads to the activation of maturation promoting factor (MPF), a heterodimer consisting of the protein kinase cdk1 and cyclin B1; this complex promotes the oocyte's entry into M phase of meiosis I. A crucial event required for MPF activation is cytoplasmic polyadenylation element (CPE)-mediated translation of specific dormant mRNAs such as c-mos and cyclin B1. The CPE, which resides in mRNA 3' untranslated region (UTR), is bound by the CPE binding protein (CPEB), which in turn is bound by Maskin. Maskin is bound to the 5' cap binding protein eIF4E. This type of closed-loop mRNA structure inhibits the recruitment and assembly of the translation initiation complex at the 5'UTR of CPE containing mRNAs. To alleviate this inhibition, CPEB undergoes phosphorylation on S174 by the serine/threonine kinase Aurora A. Phosphorylated CPEB promotes the recruitment of specific polyadenylation factors leading to the polyadenylation of the dormant mRNA, resulting in the disassociation of Maskin from eIF4E. eIF4E is subsequently bound by translation initiation factors leading to mRNA assembly into polysomes and synthesis of the encoded protein. Insulin signaling has also been shown to induce oocyte maturation. However, this signaling cascade uniquely requires the activation of two upstream components, PI3 kinase and PKC zeta. In this thesis, I show that insulin induced oocyte maturation requires the same CPE-mediated mRNA translation mechanism as had been described for progesterone signaling. I also show that Aurora A kinase activation and S174 phosphorylation play an essential role in insulin-induced CPE-mediated mRNA translation. Interestingly, inhibition of PI3 kinase and PKC zeta inhibits CPE-mediated polyadenylation only in the insulin-signaling pathway; the progesterone pathway is unaffected. These results clearly indicate that different upstream signaling components control CPE-mediated translation between progesterone and insulin signaling cascades. However, both pathways are antagonized by over expressed GSK-3, leading to inhibition of oocyte maturation. Furthermore, I found that GSK-3 inhibits Aurora A kinase activity by directly phosphorylating Aurora A on serine 290/291, promoting an inhibitory autophosphorylation event on serine 349. The importance of a GSK-3/Aurora A interaction is underscored by the finding that GSK-3, Axin, and Aurora A reside in a complex in immature oocytes. During progesterone or insulin signaling, GSK-3 dissociates from Aurora A allowing Aurora A to become active, leading to CPEB phosphorylation, CPE-mediated mRNA translation and oocyte maturation.
dc.language.isoen_US
dc.publisherUniversity of Massachusetts Medical School
dc.rightsCopyright is held by the author, with all rights reserved.
dc.subjectInsulin
dc.subjectOocytes
dc.subjectProgesterone
dc.subjectProtein Kinases
dc.subjectSignal Transduction
dc.subjectTranscription Factors
dc.subjectProtein Biosynthesis
dc.subjectXenopus laevis
dc.subjectXenopus Proteins
dc.subjectAcademic Dissertations
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleSignaling Events Leading to CPEB-Mediated Translation: a Dissertation
dc.typeDoctoral Dissertation
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_diss/305
dc.legacy.embargo2017-04-24T00:00:00-07:00
dc.identifier.contextkey294678
html.description.abstract<p>Fully grown oocytes' of the African clawed frog, <em>Xenopus laevis</em>, are arrested at the diplotene stage of meiotic prophase I, which resembles the G2 phase of the mitotic cell cycle. Re-entry into the meiotic divisions is initiated by hormonal signaling normally provided by progesterone. Progesterone signaling leads to the activation of maturation promoting factor (MPF), a heterodimer consisting of the protein kinase cdk1 and cyclin B1; this complex promotes the oocyte's entry into M phase of meiosis I. A crucial event required for MPF activation is cytoplasmic polyadenylation element (CPE)-mediated translation of specific dormant mRNAs such as c-mos and cyclin B1. The CPE, which resides in mRNA 3' untranslated region (UTR), is bound by the CPE binding protein (CPEB), which in turn is bound by Maskin. Maskin is bound to the 5' cap binding protein eIF4E. This type of closed-loop mRNA structure inhibits the recruitment and assembly of the translation initiation complex at the 5'UTR of CPE containing mRNAs. To alleviate this inhibition, CPEB undergoes phosphorylation on S174 by the serine/threonine kinase Aurora A. Phosphorylated CPEB promotes the recruitment of specific polyadenylation factors leading to the polyadenylation of the dormant mRNA, resulting in the disassociation of Maskin from eIF4E. eIF4E is subsequently bound by translation initiation factors leading to mRNA assembly into polysomes and synthesis of the encoded protein.</p> <p>Insulin signaling has also been shown to induce oocyte maturation. However, this signaling cascade uniquely requires the activation of two upstream components, PI3 kinase and PKC zeta. In this thesis, I show that insulin induced oocyte maturation requires the same CPE-mediated mRNA translation mechanism as had been described for progesterone signaling. I also show that Aurora A kinase activation and S174 phosphorylation play an essential role in insulin-induced CPE-mediated mRNA translation. Interestingly, inhibition of PI3 kinase and PKC zeta inhibits CPE-mediated polyadenylation only in the insulin-signaling pathway; the progesterone pathway is unaffected. These results clearly indicate that different upstream signaling components control CPE-mediated translation between progesterone and insulin signaling cascades. However, both pathways are antagonized by over expressed GSK-3, leading to inhibition of oocyte maturation. Furthermore, I found that GSK-3 inhibits Aurora A kinase activity by directly phosphorylating Aurora A on serine 290/291, promoting an inhibitory autophosphorylation event on serine 349. The importance of a GSK-3/Aurora A interaction is underscored by the finding that GSK-3, Axin, and Aurora A reside in a complex in immature oocytes. During progesterone or insulin signaling, GSK-3 dissociates from Aurora A allowing Aurora A to become active, leading to CPEB phosphorylation, CPE-mediated mRNA translation and oocyte maturation.</p>
dc.identifier.submissionpathgsbs_diss/305
dc.contributor.departmentGraduate School of Biomedical Sciences
dc.contributor.departmentProgram in Molecular Medicine


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