Analysis of RAD9 functions: Roles in the checkpoint response, DNA damage processing, and prevention of genomic instability

Persistent Link:
http://hdl.handle.net/10150/280312
Title:
Analysis of RAD9 functions: Roles in the checkpoint response, DNA damage processing, and prevention of genomic instability
Author:
Nyberg, Kara Ann
Issue Date:
2003
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
In the 15 years since Rad9's discovery, we have come to understand a great about Rad9 biology, yet numerous questions still remain. Especially intriguing questions include: (a) How does Rad9 get localized and/or recognize DNA damage? (b) How does Rad9 activate downstream checkpoint proteins? and (c) Does Rad9 play additional roles in recognizing and/or repairing DNA damage that have yet to be discovered? In an effort to try to answer some of these questions, I analyzed the effects of various RAD9 mutations. To assess the contribution of RAD9 to inhibiting DNA degradation and its role in the cell cycle arrest and DNA damage repair responses, I performed a pentapeptide mutagenesis screen in order to obtain RAD9 separation-of-function mutants that were proficient for one known phenotype and deficient in the other. I was able to obtain 2 such mutants that were hypomorphic in their ability to prevent DNA degradation and completely proficient for arresting the cell cycle in the presence of DNA damage and for repairing such damage. Despite many efforts, I was unable to enhance the hypomorphic dysfuntion of these mutants in preventing DNA degradation such that a suppressor screen to identify other genes in this pathway could be performed. In another effort to try to understand the role of the BRCT domains for RAD9 functions, I analyzed the effects of other various RAD9 mutants. By deletion analysis, I was able to determine that the predominant function of RAD9's BRCT domains is to mediate concentration of the Rad9 protein for function by two means: (1) by conferring stability to the Rad9 protein, and (2) by homodimerizing Rad9 to increase its local concentration to enable interactions with downstream checkpoint components.
Type:
text; Dissertation-Reproduction (electronic)
Keywords:
Biology, Molecular.; Biology, Genetics.; Biology, Cell.; Chemistry, Biochemistry.
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Graduate College; Molecular and Cellular Biology
Degree Grantor:
University of Arizona
Advisor:
Weinert, Ted A.

Full metadata record

DC FieldValue Language
dc.language.isoen_USen_US
dc.titleAnalysis of RAD9 functions: Roles in the checkpoint response, DNA damage processing, and prevention of genomic instabilityen_US
dc.creatorNyberg, Kara Annen_US
dc.contributor.authorNyberg, Kara Annen_US
dc.date.issued2003en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractIn the 15 years since Rad9's discovery, we have come to understand a great about Rad9 biology, yet numerous questions still remain. Especially intriguing questions include: (a) How does Rad9 get localized and/or recognize DNA damage? (b) How does Rad9 activate downstream checkpoint proteins? and (c) Does Rad9 play additional roles in recognizing and/or repairing DNA damage that have yet to be discovered? In an effort to try to answer some of these questions, I analyzed the effects of various RAD9 mutations. To assess the contribution of RAD9 to inhibiting DNA degradation and its role in the cell cycle arrest and DNA damage repair responses, I performed a pentapeptide mutagenesis screen in order to obtain RAD9 separation-of-function mutants that were proficient for one known phenotype and deficient in the other. I was able to obtain 2 such mutants that were hypomorphic in their ability to prevent DNA degradation and completely proficient for arresting the cell cycle in the presence of DNA damage and for repairing such damage. Despite many efforts, I was unable to enhance the hypomorphic dysfuntion of these mutants in preventing DNA degradation such that a suppressor screen to identify other genes in this pathway could be performed. In another effort to try to understand the role of the BRCT domains for RAD9 functions, I analyzed the effects of other various RAD9 mutants. By deletion analysis, I was able to determine that the predominant function of RAD9's BRCT domains is to mediate concentration of the Rad9 protein for function by two means: (1) by conferring stability to the Rad9 protein, and (2) by homodimerizing Rad9 to increase its local concentration to enable interactions with downstream checkpoint components.en_US
dc.typetexten_US
dc.typeDissertation-Reproduction (electronic)en_US
dc.subjectBiology, Molecular.en_US
dc.subjectBiology, Genetics.en_US
dc.subjectBiology, Cell.en_US
dc.subjectChemistry, Biochemistry.en_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.disciplineMolecular and Cellular Biologyen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorWeinert, Ted A.en_US
dc.identifier.proquest3089997en_US
dc.identifier.bibrecord.b44425326en_US
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