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dc.contributor.advisorGaudet, Rachelle
dc.contributor.authorWagner, Nico
dc.date.accessioned2019-12-12T08:54:23Z
dash.embargo.terms2020-05-01
dc.date.created2019-05
dc.date.issued2019-05-13
dc.date.submitted2019
dc.identifier.citationWagner, Nico. 2019. Stop Codon Readthrough in VEGF-a Is Regulated by Synergistic Activity of a Complex Tripartite Signal. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences.
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:42029674*
dc.description.abstractTranslational recoding via frameshifting or stop-codon readthrough is generally regulated by a structured mRNA signal downstream of the slippery sequence or stop-codon, respectively. The strength of this signal can be enhanced or attenuated by neighboring sequences, and in some rare cases, the signal is known to interact with sequences hundreds of nucleotides downstream in the 3’ region to regulate the level of recoding. In contrast, here we show that recoding of the VEGF-A mRNA is regulated by three distinct elements that can each independently support stop codon readthrough: one, the recently discovered Ax-element downstream of the stop codon; and two other elements, Au1 and Au2 in the coding region ~500 and ~250 nucleotides upstream of the stop codon, respectively. The Ax-element folds into three stem loops (SL-Ax1-3) while the Au elements are predicted to fold into single stem loops. Furthermore, we also show that the trans-acting protein hnRNP A2/B1 independently interacts with SL-Au1 and SL-Ax1 to confer efficient readthrough. While the Ax-element individually allows for only 0.2% readthrough, and each Au element for 0.25% readthrough levels, together they allow for a robust 2% readthrough level. Given that structural manipulations of SL-Au1 at the 5’ end the coding sequences led to a 16% readthrough activity of the stop codon at the 3’ end, the observed synergistic activity is potentially indicative of long-range interactions. Our study thus highlights both the control of absolute readthrough levels through strategic structures and the functional importance of maintenance of 5’ - 3’ end interactions during the process of active translation.
dc.description.sponsorshipBiology, Molecular and Cellular
dc.format.mimetypeapplication/pdf
dc.language.isoen
dash.licenseLAA
dc.subjectVEGF-A
dc.subjectreadthrough
dc.subjectrecoding
dc.subjecthnRNP A2/B1
dc.titleStop Codon Readthrough in VEGF-a Is Regulated by Synergistic Activity of a Complex Tripartite Signal
dc.typeThesis or Dissertation
dash.depositing.authorWagner, Nico
dash.embargo.until2020-05-01
dc.date.available2019-12-12T08:54:23Z
thesis.degree.date2019
thesis.degree.grantorGraduate School of Arts & Sciences
thesis.degree.grantorGraduate School of Arts & Sciences
thesis.degree.levelDoctoral
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy
thesis.degree.nameDoctor of Philosophy
dc.contributor.committeeMemberGarner, Ethan
dc.contributor.committeeMemberWagner, Gerhard
dc.type.materialtext
thesis.degree.departmentBiology, Molecular and Cellular
thesis.degree.departmentBiology, Molecular and Cellular
dash.identifier.vireo
dc.identifier.orcid0000-0001-8200-3878
dash.author.emailwagnernico@gmail.com


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