Amyloid precursor protein translation is regulated by a 3'UTR guanine quadruplex

Ezekiel Crenshaw, Brian P. Leung, Chun Kit Kwok, Michal Sharoni, Kalee Olson, Neeraj P. Sebastian, Sara Ansaloni, Reinhard Schweitzer-Stenner, Michael R. Akins, Philip C. Bevilacqua, Aleister J. Saunders

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A central event in Alzheimer's disease is the accumulation of amyloid β (Aβ) peptides generated by the proteolytic cleavage of the amyloid precursor protein (APP). APP overexpression leads to increased Aβ generation and Alzheimer's disease in humans and altered neuronal migration and increased long term depression in mice. Conversely, reduction of APP expression results in decreased Aβ levels in mice as well as impaired learning and memory and decreased numbers of dendritic spines. Together these findings indicate that therapeutic interventions that aim to restore APP and Aβ levels must do so within an ideal range. To better understand the effects of modulating APP levels, we explored the mechanisms regulating APP expression focusing on post-transcriptional regulation. Such regulation can be mediated by RNA regulatory elements such as guanine quadruplexes (Gquadruplexes), non-canonical structured RNA motifs that affect RNA stability and translation. Via a bioinformatics approach, we identified a candidate G-quadruplex within the APP mRNA in its 3'UTR (untranslated region) at residues 3008-3027 (NM201414.2). This sequence exhibited characteristics of a parallel G-quadruplex structure as revealed by circular dichroism spectrophotometry. Further, as with other G-quadruplexes, the formation of this structure was dependent on the presence of potassium ions. This G-quadruplex has no apparent role in regulating transcription or mRNA stability as wild type and mutant constructs exhibited equivalent mRNA levels as determined by real time PCR. Instead, we demonstrate that this G-quadruplex negatively regulates APP protein expression using dual luciferase reporter and Western blot analysis. Taken together, our studies reveal post-transcriptional regulation by a 3'UTR G-quadruplex as a novel mechanism regulating APP expression.

Original languageEnglish (US)
Article numbere0143160
JournalPloS one
Volume10
Issue number11
DOIs
StatePublished - Nov 1 2015

Fingerprint

G-Quadruplexes
Amyloid beta-Protein Precursor
guanine
Guanine
Protein Biosynthesis
3' Untranslated Regions
3' untranslated regions
amyloid
Amyloid
translation (genetics)
proteins
protein synthesis
Serum Amyloid A Protein
RNA
RNA Stability
Messenger RNA
Alzheimer disease
Alzheimer Disease
Untranslated Regions
Dendritic Spines

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Crenshaw, E., Leung, B. P., Kwok, C. K., Sharoni, M., Olson, K., Sebastian, N. P., ... Saunders, A. J. (2015). Amyloid precursor protein translation is regulated by a 3'UTR guanine quadruplex. PloS one, 10(11), [e0143160]. https://doi.org/10.1371/journal.pone.0143160
Crenshaw, Ezekiel ; Leung, Brian P. ; Kwok, Chun Kit ; Sharoni, Michal ; Olson, Kalee ; Sebastian, Neeraj P. ; Ansaloni, Sara ; Schweitzer-Stenner, Reinhard ; Akins, Michael R. ; Bevilacqua, Philip C. ; Saunders, Aleister J. / Amyloid precursor protein translation is regulated by a 3'UTR guanine quadruplex. In: PloS one. 2015 ; Vol. 10, No. 11.
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abstract = "A central event in Alzheimer's disease is the accumulation of amyloid β (Aβ) peptides generated by the proteolytic cleavage of the amyloid precursor protein (APP). APP overexpression leads to increased Aβ generation and Alzheimer's disease in humans and altered neuronal migration and increased long term depression in mice. Conversely, reduction of APP expression results in decreased Aβ levels in mice as well as impaired learning and memory and decreased numbers of dendritic spines. Together these findings indicate that therapeutic interventions that aim to restore APP and Aβ levels must do so within an ideal range. To better understand the effects of modulating APP levels, we explored the mechanisms regulating APP expression focusing on post-transcriptional regulation. Such regulation can be mediated by RNA regulatory elements such as guanine quadruplexes (Gquadruplexes), non-canonical structured RNA motifs that affect RNA stability and translation. Via a bioinformatics approach, we identified a candidate G-quadruplex within the APP mRNA in its 3'UTR (untranslated region) at residues 3008-3027 (NM201414.2). This sequence exhibited characteristics of a parallel G-quadruplex structure as revealed by circular dichroism spectrophotometry. Further, as with other G-quadruplexes, the formation of this structure was dependent on the presence of potassium ions. This G-quadruplex has no apparent role in regulating transcription or mRNA stability as wild type and mutant constructs exhibited equivalent mRNA levels as determined by real time PCR. Instead, we demonstrate that this G-quadruplex negatively regulates APP protein expression using dual luciferase reporter and Western blot analysis. Taken together, our studies reveal post-transcriptional regulation by a 3'UTR G-quadruplex as a novel mechanism regulating APP expression.",
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Crenshaw, E, Leung, BP, Kwok, CK, Sharoni, M, Olson, K, Sebastian, NP, Ansaloni, S, Schweitzer-Stenner, R, Akins, MR, Bevilacqua, PC & Saunders, AJ 2015, 'Amyloid precursor protein translation is regulated by a 3'UTR guanine quadruplex', PloS one, vol. 10, no. 11, e0143160. https://doi.org/10.1371/journal.pone.0143160

Amyloid precursor protein translation is regulated by a 3'UTR guanine quadruplex. / Crenshaw, Ezekiel; Leung, Brian P.; Kwok, Chun Kit; Sharoni, Michal; Olson, Kalee; Sebastian, Neeraj P.; Ansaloni, Sara; Schweitzer-Stenner, Reinhard; Akins, Michael R.; Bevilacqua, Philip C.; Saunders, Aleister J.

In: PloS one, Vol. 10, No. 11, e0143160, 01.11.2015.

Research output: Contribution to journalArticle

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AU - Crenshaw, Ezekiel

AU - Leung, Brian P.

AU - Kwok, Chun Kit

AU - Sharoni, Michal

AU - Olson, Kalee

AU - Sebastian, Neeraj P.

AU - Ansaloni, Sara

AU - Schweitzer-Stenner, Reinhard

AU - Akins, Michael R.

AU - Bevilacqua, Philip C.

AU - Saunders, Aleister J.

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N2 - A central event in Alzheimer's disease is the accumulation of amyloid β (Aβ) peptides generated by the proteolytic cleavage of the amyloid precursor protein (APP). APP overexpression leads to increased Aβ generation and Alzheimer's disease in humans and altered neuronal migration and increased long term depression in mice. Conversely, reduction of APP expression results in decreased Aβ levels in mice as well as impaired learning and memory and decreased numbers of dendritic spines. Together these findings indicate that therapeutic interventions that aim to restore APP and Aβ levels must do so within an ideal range. To better understand the effects of modulating APP levels, we explored the mechanisms regulating APP expression focusing on post-transcriptional regulation. Such regulation can be mediated by RNA regulatory elements such as guanine quadruplexes (Gquadruplexes), non-canonical structured RNA motifs that affect RNA stability and translation. Via a bioinformatics approach, we identified a candidate G-quadruplex within the APP mRNA in its 3'UTR (untranslated region) at residues 3008-3027 (NM201414.2). This sequence exhibited characteristics of a parallel G-quadruplex structure as revealed by circular dichroism spectrophotometry. Further, as with other G-quadruplexes, the formation of this structure was dependent on the presence of potassium ions. This G-quadruplex has no apparent role in regulating transcription or mRNA stability as wild type and mutant constructs exhibited equivalent mRNA levels as determined by real time PCR. Instead, we demonstrate that this G-quadruplex negatively regulates APP protein expression using dual luciferase reporter and Western blot analysis. Taken together, our studies reveal post-transcriptional regulation by a 3'UTR G-quadruplex as a novel mechanism regulating APP expression.

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Crenshaw E, Leung BP, Kwok CK, Sharoni M, Olson K, Sebastian NP et al. Amyloid precursor protein translation is regulated by a 3'UTR guanine quadruplex. PloS one. 2015 Nov 1;10(11). e0143160. https://doi.org/10.1371/journal.pone.0143160