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Isolation and characterization of lipid rafts in Emiliania huxley

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TitleInfo
Title
Isolation and characterization of lipid rafts in Emiliania huxley
SubTitle
key players in host-virus interactions
Name (type = personal)
NamePart (type = family)
Rose
NamePart (type = given)
Suzanne L.
NamePart (type = date)
1968-
DisplayForm
Suzanne Rose
Role
RoleTerm (authority = RULIB)
author
Name (type = personal)
NamePart (type = family)
Bidle
NamePart (type = given)
Kay D
DisplayForm
Kay D Bidle
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
chair
Name (type = personal)
NamePart (type = family)
Schofield
NamePart (type = given)
Oscar
DisplayForm
Oscar Schofield
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
internal member
Name (type = personal)
NamePart (type = family)
Duffy
NamePart (type = given)
Sioban
DisplayForm
Sioban Duffy
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
internal member
Name (type = personal)
NamePart (type = family)
Bhattacharya
NamePart (type = given)
Debashish
DisplayForm
Debashish Bhattacharya
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
outside member
Name (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB)
degree grantor
Name (type = corporate)
NamePart
Graduate School - New Brunswick
Role
RoleTerm (authority = RULIB)
school
TypeOfResource
Text
Genre (authority = marcgt)
theses
OriginInfo
DateCreated (qualifier = exact)
2013
DateOther (qualifier = exact); (type = degree)
2013-01
Place
PlaceTerm (type = code)
xx
Language
LanguageTerm (authority = ISO639-2b); (type = code)
eng
Abstract (type = abstract)
Coccolithoviruses (EhVs) employ a suite of glycosphingolipids (GSLs) to successfully infect the globally important coccolithophore Emiliania huxleyi. Given their enrichment in glycosphingolipids and their involvement in sensing extracellular stimuli and activating signaling cascades through protein-protein interactions, lipid rafts and associated microdomains likely play a heretofore unappreciated role in viral infectivity and resistance mechanisms. Indeed, the lipid raft protein flotillin carries an evolutionary conserved prohibitin homology (PHB) domain, which has been shown to influence pathogen resistance by inducing hypersensitive response and cell death. Little to nothing is known about lipid rafts in unicellular photoautotrophic organisms, especially those from the marine environment. We combined bioinformatics, detergent treatment, density gradient purification, western blotting, lipidomics and genome-enabled proteomics to isolate and characterize the lipid and protein content of lipid rafts from control and EhV86-infected E. huxleyi strain CCMP1516. Using bioinformatic analysis, we identified a flotillin-like protein (Protein ID 363433) in the E. huxleyi CCMP1516 genome, which contained several evolutionary-conserved features of known flotillin proteins. Western blot analysis further revealed immunoreactivity of a 67 kDa protein in E. huxleyi cell extracts to a human anti-flotillin antibody, consistent with the dimerized form of this protein and thereby providing a biochemical marker for lipid raft isolation. Lipid raft-enriched fractions were isolated and purified as detergent (Brij96) resistant membranes (DRMs) in Optiprep density gradients and their lipid and protein composition were subsequently characterized. TSQ mass spectrometry of the lipids in DRM fractions showed novel host-derived GSLs which co-purified with flotillin immunoreactive fractions. These fractions also contained notable vesicle morphology as confirmed by TEM. Subsequent analysis of lipid raft proteome in both uninfected and EhV86-infected E. huxleyi cells collected at 2 h confirmed flotillin as one of the major proteins and more broadly provided key insight into host defense and virus infection strategies at the interface of lipid rafts and stress, programmed cell death, and innate immunity pathways. In particular, the detection of an EhV86-encoded C-type lectin (CTL)-containing protein (ehv149), with putative roles in cell adhesion and pathogen recognition receptors, suggests that EhV86 infection of E. huxleyi occurs by way of the recognition of specific glycosphingolipids and raft-associated proteins.
Subject (authority = RUETD)
Topic
Ecology and Evolution
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_4436
PhysicalDescription
Form (authority = gmd)
electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
viii, 47 p. : ill.
Note (type = degree)
M.S.
Note (type = bibliography)
Includes bibliographical references
Note (type = statement of responsibility)
by Suzanne L. Rose
Subject (authority = ETD-LCSH)
Topic
Coccolithus huxleyi--Virus diseases
Subject (authority = ETD-LCSH)
Topic
Lipids--Research
Identifier (type = hdl)
http://hdl.rutgers.edu/1782.1/rucore10001600001.ETD.000067826
RelatedItem (type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier (type = local)
rucore19991600001
Location
PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier (type = doi)
doi:10.7282/T3D50KN3
Genre (authority = ExL-Esploro)
ETD graduate
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Rights

RightsDeclaration (ID = rulibRdec0006)
The author owns the copyright to this work.
RightsHolder (type = personal)
Name
FamilyName
Rose
GivenName
Suzanne
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime (encoding = w3cdtf); (qualifier = exact); (point = start)
2012-12-19 10:58:36
AssociatedEntity
Name
Suzanne Rose
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
AssociatedObject
Type
License
Name
Author Agreement License
Detail
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.
Copyright
Status
Copyright protected
Availability
Status
Open
Reason
Permission or license
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Technical

RULTechMD (ID = TECHNICAL1)
ContentModel
ETD
OperatingSystem (VERSION = 5.1)
windows xp
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