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Phosphorylation of lipin 1 beta phosphatidate phosphatase by casein kinase II
Descriptive
TitleInfo
Title
Phosphorylation of lipin 1 beta phosphatidate phosphatase by casein kinase II
Name
(type = personal)
NamePart
(type = family)
Hennessy
NamePart
(type = given)
Meagan L.
NamePart
(type = date)
1980-
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Meagan L. Hennessy
Role
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(authority = RULIB)
author
Name
(type = personal)
NamePart
(type = family)
Carman
NamePart
(type = given)
George M
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George M Carman
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Advisory Committee
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chair
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(type = personal)
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Storch
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(type = given)
Judith
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Judith Storch
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Advisory Committee
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(authority = RULIB)
internal member
Name
(type = personal)
NamePart
(type = family)
Watford
NamePart
(type = given)
Malcolm
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Malcolm Watford
Affiliation
Advisory Committee
Role
RoleTerm
(authority = RULIB)
internal member
Name
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NamePart
Rutgers University
Role
RoleTerm
(authority = RULIB)
degree grantor
Name
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NamePart
School of Graduate Studies
Role
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(authority = RULIB)
school
TypeOfResource
Text
Genre
(authority = marcgt)
theses
OriginInfo
DateCreated
(qualifier = exact)
2018
DateOther
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(type = degree)
2018-10
CopyrightDate
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2018
Place
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(type = code)
xx
Language
LanguageTerm
(authority = ISO639-2b);
(type = code)
eng
Abstract
(type = abstract)
Lipin is a phosphatidate phosphatase enzyme that catalyzes the penultimate step in triacylglycerol synthesis, the conversion of phosphatidic acid to diacylglycerol. By controlling this step, lipin regulates both triacylglycerol and phospholipid synthesis, and it also acts as a transcriptional regulator. It is known that phosphorylation controls the subcellular localization of lipin, and its activity as both a phosphatidate phosphatase and a transcriptional regulator. Multiple phosphorylation sites within lipin have been identified, but the identity of the protein kinases involved is largely unknown. In this work, we sought to identify and characterize kinases that play a role in lipin’s regulation.
Using bioinformatics, we found that lipin 1 beta has a high probability of being phosphorylated by casein kinase II. Using purified lipin 1 beta, we showed that casein kinase II catalyzed the incorporation of the gamma-phosphate of [gamma-32P]ATP into the protein. We then focused on the serine-rich domain of lipin 1 beta, which in lipin 1 beta has been shown to be important in subcellular localization through interaction with 14-3-3 protein. We generated a 22-residue peptide that included 7 putative serine/threonine phosphorylation sites, and found that this peptide was phosphorylated by casein kinase II. Phosphorylation analysis of the peptide with alanine substitutions for the putative sites indicated that Ser-285 and Ser-287 were the targets of the kinase. In full-length lipin 1 beta, the S285A mutation caused a greater degree of phosphorylation, whereas the extent of phosphorylation of the S285A/S287A mutant was about the same as that of the control wild type protein. In phosphopeptide mapping analysis, two phosphopeptides present in the wild type protein were absent in both mutant proteins, but one phosphopeptide present in the mutants was absent in the wild type protein. These observations indicate that the phosphorylation by casein kinase II is complex. This work increases our understanding of the posttranslational modification of lipin 1 beta by phosphorylation.
Using bioinformatics, we found that lipin 1 beta has a high probability of being phosphorylated by casein kinase II. Using purified lipin 1 beta, we showed that casein kinase II catalyzed the incorporation of the gamma-phosphate of [gamma-32P]ATP into the protein. We then focused on the serine-rich domain of lipin 1 beta, which in lipin 1 beta has been shown to be important in subcellular localization through interaction with 14-3-3 protein. We generated a 22-residue peptide that included 7 putative serine/threonine phosphorylation sites, and found that this peptide was phosphorylated by casein kinase II. Phosphorylation analysis of the peptide with alanine substitutions for the putative sites indicated that Ser-285 and Ser-287 were the targets of the kinase. In full-length lipin 1 beta, the S285A mutation caused a greater degree of phosphorylation, whereas the extent of phosphorylation of the S285A/S287A mutant was about the same as that of the control wild type protein. In phosphopeptide mapping analysis, two phosphopeptides present in the wild type protein were absent in both mutant proteins, but one phosphopeptide present in the mutants was absent in the wild type protein. These observations indicate that the phosphorylation by casein kinase II is complex. This work increases our understanding of the posttranslational modification of lipin 1 beta by phosphorylation.
Subject
(authority = RUETD)
Topic
Nutritional Sciences
Subject
(authority = ETD-LCSH)
Topic
Bioinformatics
Subject
(authority = ETD-LCSH)
Topic
Phosphorylation
Subject
(authority = local)
Topic
Lipin
RelatedItem
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Title
Rutgers University Electronic Theses and Dissertations
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ETD
Identifier
ETD_9155
PhysicalDescription
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electronic resource
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application/pdf
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text/xml
Extent
1 online resource (83 pages) : illustrations
Note
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M.S.
Note
(type = bibliography)
Includes bibliographical references
Note
(type = statement of responsibility)
by Meagan L. Hennessy
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TitleInfo
Title
School of Graduate Studies Electronic Theses and Dissertations
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rucore10001600001
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NjNbRU
Identifier
(type = doi)
doi:10.7282/t3-atsq-5520
Genre
(authority = ExL-Esploro)
ETD graduate
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Rights
RightsDeclaration
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The author owns the copyright to this work.
RightsHolder
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Name
FamilyName
Hennessy
GivenName
Meagan
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime
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(qualifier = exact);
(point = start)
2018-08-22 15:19:14
AssociatedEntity
Name
Meagan Hennessy
Role
Copyright holder
Affiliation
Rutgers University. School of Graduate Studies
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License
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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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Embargo
DateTime
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2018-10-31
DateTime
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2019-10-31
Detail
Access to this PDF has been restricted at the author's request. It will be publicly available after October 31st, 2019.
Copyright
Status
Copyright protected
Availability
Status
Open
Reason
Permission or license
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windows xp
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2018-09-28T02:23:59
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2018-09-28T02:23:59
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