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Structural characterization and transcriptional regulation of the cytosolic PSD-95 interacting protein (cypin) and its role in neuronal dendrite branching
Descriptive
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Title
Structural characterization and transcriptional regulation of the cytosolic PSD-95 interacting protein
(cypin) and its role in neuronal dendrite branching
(cypin) and its role in neuronal dendrite branching
Name
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Fernández
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José R.
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José R. Fernández
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author
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Firestein
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Bonnie
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Advisory Committee
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Bonnie L Firestein
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chair
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Welsh
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William
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Advisory Committee
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William J Welsh
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internal member
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Brewer
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Gary
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Advisory Committee
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Gary Brewer
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internal member
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Byrne
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Bruce
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Advisory Committee
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Bruce C Byrne
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outside member
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Rutgers University
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degree grantor
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Graduate School - New Brunswick
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Text
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theses
OriginInfo
DateCreated
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2008
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2008-10
Language
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English
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electronic
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application/pdf
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text/xml
Extent
xii, 172 pages
Abstract
Dendrite morphology regulates how a postsynaptic neuron receives information from presynaptic neurons. The specific patterning of dendrite branches is promoted by extrinsic and intrinsic factors that trigger the activation of functional signaling pathways. However, only a handful of the regulatory factors and biochemical mechanisms involved in determining dendrite morphology are known. The Firestein laboratory previously reported that cypin (cytosolic PSD-95 interactor), the mammalian guanine deaminase (GDA), plays an active role in regulating dendrite branching in hippocampal neurons. Cypin-promoted increases in dendrite number are dependent on binding of zinc ions to cypin and cypin's guanine deaminase activity.
This work focuses on the identification of specific structural properties of cypin that lead to the multifunctional roles in guanine metabolism and dendrite development. We first employed phylogenetic analysis and computational structure modeling techniques to construct a three dimensional structural model of cypin. Inspection of our structural model confirmed that specific predicted residues coordinate with a zinc ion to play a role in enzymatic activity and promotion of dendrite branching in developing neurons. In addition, we used a combination of protein structure analysis, experimental kinetic studies, and cell culture tests to uncover novel potential ligands for cypin. We obtained a list of compounds that demonstrate higher binding affinity to GDA than does guanine. Our results provide evidence that an in silico drug discovery strategy coupled with in vitro verification can be successfully implemented to discover compounds that may have therapeutic value for the treatment of diseases and disorders where GDA activity is abnormal.
Since the regulation of dendrite branching function by cypin is dependent on intracellular levels of cypin protein, idenitification of transcriptional regulators of cypin gene expression may elucidate how cypin regulates neurite development. Therefore, we also investigated how extracellular factors can regulate cypin expression. Our data show that in developing neurons, BDNF increases cypin protein via activation of the MEK pathway, and consequently, CREB transcription factor-dependent cypin gene expression. The discovery of intrinsic regulators of cypin expression aids in our understanding of molecular mechanisms underlying dendritic patterning, and hence, synaptic plasticity, learning and memory.
This work focuses on the identification of specific structural properties of cypin that lead to the multifunctional roles in guanine metabolism and dendrite development. We first employed phylogenetic analysis and computational structure modeling techniques to construct a three dimensional structural model of cypin. Inspection of our structural model confirmed that specific predicted residues coordinate with a zinc ion to play a role in enzymatic activity and promotion of dendrite branching in developing neurons. In addition, we used a combination of protein structure analysis, experimental kinetic studies, and cell culture tests to uncover novel potential ligands for cypin. We obtained a list of compounds that demonstrate higher binding affinity to GDA than does guanine. Our results provide evidence that an in silico drug discovery strategy coupled with in vitro verification can be successfully implemented to discover compounds that may have therapeutic value for the treatment of diseases and disorders where GDA activity is abnormal.
Since the regulation of dendrite branching function by cypin is dependent on intracellular levels of cypin protein, idenitification of transcriptional regulators of cypin gene expression may elucidate how cypin regulates neurite development. Therefore, we also investigated how extracellular factors can regulate cypin expression. Our data show that in developing neurons, BDNF increases cypin protein via activation of the MEK pathway, and consequently, CREB transcription factor-dependent cypin gene expression. The discovery of intrinsic regulators of cypin expression aids in our understanding of molecular mechanisms underlying dendritic patterning, and hence, synaptic plasticity, learning and memory.
Note
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Ph.D.
Note
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Includes bibliographical references (p. 145-171).
Subject
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Topic
Pharmacology, Cellular and Molecular
Subject
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Topic
Neurons--Physiology
Subject
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Topic
Nervous system
Subject
(ID = SUBJ4);
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Topic
Dendrites
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Title
Graduate School - New Brunswick Electronic Theses and Dissertations
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rucore19991600001
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http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.17468
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ETD_1124
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doi:10.7282/T36H4HRQ
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ETD doctoral
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The author owns the copyright to this work.
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Copyright protected
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Open
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Name
Jos� Fern�ndez
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Rutgers University. Graduate School - New Brunswick
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Non-exclusive ETD license
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I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
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