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Regulation of expression of copper responsive genes in Sulfolobus solfataricus

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TypeOfResource
Text
TitleInfo (ID = T-1)
Title
Regulation of expression of copper responsive genes in Sulfolobus solfataricus
SubTitle
PartName
PartNumber
NonSort
Identifier (displayLabel = ); (invalid = )
ETD_2387
Identifier (type = hdl)
http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.000052159
Language (objectPart = )
LanguageTerm (authority = ISO639-2); (type = code)
eng
Genre (authority = marcgt)
theses
Subject (ID = SBJ-1); (authority = RUETD)
Topic
Microbiology and Molecular Genetics
Subject (ID = SBJ-2); (authority = ETD-LCSH)
Topic
Genetic regulation
Subject (ID = SBJ-3); (authority = ETD-LCSH)
Topic
Copper in the body
Abstract
Copper is an essential micronutrient, but toxic in excess. Cells must maintain their internal level of copper within a narrow range of concentrations. This is accomplished mainly by copper efflux mediated by ATP-driven copper transporters that are induced at the level of transcription. Sulfolobus solfataricus has the ability to adapt to fluctuations of copper levels in its environment. Two Sulfolobus strains P2 and 98/2, showed different sensitivity to copper To better understand the molecular mechanism behind the organismal response to copper, the expression of the cluster of genes copRTA, which encodes the copper-responsive transcriptional regulator CopR, the copper-binding protein CopT, and CopA, has been investigated. The expression of the copR and copA transcripts was monitored by quantitative real-time RT-PCR. The data showed that only copA was induced by copper. By comparing the patterns of copA expression and cellular copper accumulation, as determined by Inductively Coupled Plasma Optical Emission spectrometry, it was concluded that the level of copA depends on the internal fluctuations of copper. To investigate the role of CopR, a mutant carrying a disruption of the copR gene was created. The mutant strain was uncapable of growth in the presence of excess copper, and under the same conditions no transcription of copA was observed. These data suggested that CopR positively regulates transcription of copA. The knockout mutant strain PBL2070, its parent strain and the mutant complemented with a wild type copy of copR, were compared with respect to their physiological and transcriptional response to copper. Results confirmed that, under copper stress, CopR, activates the transcription of copA, and its presence restores the wild type phenotype resistant to copper. Finally, a reporter system based on the thermostable β-glucuronidase of S. solfataricus was developed to study the promoter of regulated genes. This system was applied to the analysis of the copTA promoter region fused to the S. solfataricus β-glucuronidase reporter. GUS activity assays showed that after copper addition, the activity of GusB was consistent with the transcriptional changes observed for copA under similar conditions. The results provide the basis for a model of the molecular mechanisms of copper homeostasis in Sulfolobus.
PhysicalDescription
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electronic resource
Extent
xiii, 115 p. : ill.
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application/pdf
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text/xml
Note (type = degree)
Ph.D.
Note (type = bibliography)
Includes bibliographical references (p. 87-113)
Note (type = statement of responsibility)
by Aramis Alexis Villafañe Martinez
Name (ID = NAME-1); (type = personal)
NamePart (type = family)
Villafañe Martinez
NamePart (type = given)
Aramis Alexis
NamePart (type = termsOfAddress)
NamePart (type = date)
1980-
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RoleTerm (authority = RULIB); (type = )
author
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Aramis Alexis Villafañe Martinez
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Bini
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Elisabetta
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chair
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Advisory Committee
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Elisabetta Bini
Name (ID = NAME-3); (type = personal)
NamePart (type = family)
Vetriani
NamePart (type = given)
Costantino
Role
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internal member
Affiliation
Advisory Committee
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Costantino Vetriani
Name (ID = NAME-4); (type = personal)
NamePart (type = family)
Zylstra
NamePart (type = given)
Gerben
Role
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internal member
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Advisory Committee
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Gerben J Zylstra
Name (ID = NAME-5); (type = personal)
NamePart (type = family)
Montville
NamePart (type = given)
Thomas
Role
RoleTerm (authority = RULIB); (type = )
outside member
Affiliation
Advisory Committee
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Thomas J Montville
Name (ID = NAME-1); (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB); (type = )
degree grantor
Name (ID = NAME-2); (type = corporate)
NamePart
Graduate School - New Brunswick
Role
RoleTerm (authority = RULIB); (type = )
school
OriginInfo
DateCreated (point = ); (qualifier = exact)
2010
DateOther (qualifier = exact); (type = degree)
2010-01
Place
PlaceTerm (type = code)
xx
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
RelatedItem (type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier (type = local)
rucore19991600001
Location
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NjNbRU
Identifier (type = doi)
doi:10.7282/T3C53M0T
Genre (authority = ExL-Esploro)
ETD doctoral
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Rights

RightsDeclaration (AUTHORITY = GS); (ID = rulibRdec0006)
The author owns the copyright to this work.
Copyright
Status
Copyright protected
Notice
Note
Availability
Status
Open
Reason
Permission or license
Note
RightsHolder (ID = PRH-1); (type = personal)
Name
FamilyName
Villafane
GivenName
Aramis
Role
Copyright Holder
RightsEvent (ID = RE-1); (AUTHORITY = rulib)
Type
Permission or license
Label
Place
DateTime
2010-01-05 11:52:29
Detail
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Role
Copyright holder
Name
Aramis Villafane
Affiliation
Rutgers University. Graduate School - New Brunswick
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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.
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Technical

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ETD
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application/pdf
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993280
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