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Title: Construction of mitochondrion-targeted telomerase for analysis in Saccharomyces cerevisiae.
Creator: Martin, Ricardo., Harriet L. Wilkes Honors College
Abstract/Description: Telomerase is associated with telomere production and nDNA protection. However, studies by Santos et al. have demonstrated that human telomerase has a mitochondrial entry sequence and in the presence of hydrogen peroxide it has been found inside the mitochondrion and may cause mitochondrial DNA mutations. Saccharomyces cerevisiae contains telomerase, but it does not have the mitochondrial entry sequence. To determine if the presence of telomerase in the mitochondria can induce mutations an...
Show moreTelomerase is associated with telomere production and nDNA protection. However, studies by Santos et al. have demonstrated that human telomerase has a mitochondrial entry sequence and in the presence of hydrogen peroxide it has been found inside the mitochondrion and may cause mitochondrial DNA mutations. Saccharomyces cerevisiae contains telomerase, but it does not have the mitochondrial entry sequence. To determine if the presence of telomerase in the mitochondria can induce mutations an experiment was developed in which a mitochondrion entry sequence would be fused to the S. cerevisiae telomerase enzyme. This fusion could then be screened in S. cerevisiae with an ade2 mutation for a simple color assay of mitochondrial activity. To date, no successful transformant has been identified. The frequency of incorrect ligations has been recognized and may indicate that the desired fusion is lethal to E. coli cells.
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Date Issued: 2009
PURL: http://purl.flvc.org/FAU/209994
Subject Headings: Cell membranes, Formation, Mitochondrial DNA, Mutation (Biology), Cell metabolism
Format: Document (PDF)

Title: Cytogenetic of chromosomal synteny evaluation: bioinformatic applications towards screening of chromosomal aberrations/ genetic disorder.
Creator: Sharma, Sandhya, Neelakanta, Perambur S., Florida Atlantic University, College of Engineering and Computer Science, Department of Computer and Electrical Engineering and Computer Science
Abstract/Description: The research efforts refer to tracking homologus loci in the chromosomes of a pair of a species. The purpose is to infer the extent of maximum syntenic correlation when an exhaustive set of orthologs of the species are searched. Relevant bioinformatic analyses use comparative mapping of conserved synteny via Oxford grid. In medical diagnostic efforts, deducing such synteny correlation can help screening chromosomal aberration in genetic disorder pathology. Objectively, the present study...
Show moreThe research efforts refer to tracking homologus loci in the chromosomes of a pair of a species. The purpose is to infer the extent of maximum syntenic correlation when an exhaustive set of orthologs of the species are searched. Relevant bioinformatic analyses use comparative mapping of conserved synteny via Oxford grid. In medical diagnostic efforts, deducing such synteny correlation can help screening chromosomal aberration in genetic disorder pathology. Objectively, the present study addresses: (i) Cytogenetic framework of syntenic correlation and, (ii) applying information-theoretics to determine entropy-dictated synteny across an exhaustive set of orthologs of the test pairs of species.
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Date Issued: 2014
PURL: http://purl.flvc.org/fau/fd/FA00004331, http://purl.flvc.org/fau/fd/FA00004331
Subject Headings: Cytogenetics, Genetic screening, Human chromosome abnormalities, Medical genetics, Molecular biology, Molecular diagnosis, Molecular genetics, Mutation (Biology)
Format: Document (PDF)

Title: Creation of an aconitase overexpression strain of Saccharomyces cerevisiae for lifespan analysis.
Creator: Nunes, Steve., Harriet L. Wilkes Honors College
Abstract/Description: In my thesis work, I attempted to construct a plasmid that would allow stable integration of genes into the Saccharomyces cerevisiae yeast genome under the control of the repressible TetO promoter. The yeast ACO1 gene was cloned under the control of the TetO operator and the tTA transactivator. This construct was inserted into yeast cells in order to observe the effects of aconitase overexpression on aging. Unfortunately, the transformed cells appeared incapable of aconitase expression as...
Show moreIn my thesis work, I attempted to construct a plasmid that would allow stable integration of genes into the Saccharomyces cerevisiae yeast genome under the control of the repressible TetO promoter. The yeast ACO1 gene was cloned under the control of the TetO operator and the tTA transactivator. This construct was inserted into yeast cells in order to observe the effects of aconitase overexpression on aging. Unfortunately, the transformed cells appeared incapable of aconitase expression as determined by glutamic acid auxptrophy, a phenotype of aconitase mutants. We have sequenced the pIT1ACO1 plasmid and have found many abnormalities in the promoter region. If the plasmid can be made to function as intended, the resulting yeast strain can be used in the future to determine if aconitase plays an important role in cellular aging.
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Date Issued: 2012
PURL: http://purl.flvc.org/FAU/3359306
Subject Headings: Yeast fungi, Research, Methodology, Microbial genetics, Genetic engineering, Aging, Molecular aspects, Cell metabolism, Mutation (Biology)
Format: Document (PDF)

Title: The effect of mutated aconitase on yeast longevity.
Creator: Kwan, CJ., Harriet L. Wilkes Honors College
Abstract/Description: Aconitase is an important enzyme in the citric Acid Cycle, is needed for maintenance of mitochondrial DNA, is a key regulator of iron in the cell, and is very sensitive to oxidative stress. We have isolatd the yeast ACO1 gene, which codes for aconitase, and randomly mutated it to create a mutant library of cells each expressing a different version of ACO1. We will select for oxidative stress resistant aconitase in S. cerevisiae by subjecting strains to successive rounds of heat shock and...
Show moreAconitase is an important enzyme in the citric Acid Cycle, is needed for maintenance of mitochondrial DNA, is a key regulator of iron in the cell, and is very sensitive to oxidative stress. We have isolatd the yeast ACO1 gene, which codes for aconitase, and randomly mutated it to create a mutant library of cells each expressing a different version of ACO1. We will select for oxidative stress resistant aconitase in S. cerevisiae by subjecting strains to successive rounds of heat shock and competitive growth against other mutants. The "winner" of this competition will then be analyzed for which version of aconitase it is expressing, which may lead to increased longevity.
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Date Issued: 2012
PURL: http://purl.flvc.org/FAU/3359310
Subject Headings: Yeast fungi, Research, Microbial genetics, Aging, Molecular aspects, Mutation (Biology), Cell metabolism
Format: Document (PDF)

Title: Cytogenic bioinformatics of chromosomal aberrations and genetic disorders: data-mining of relevant biostatistical features.
Creator: Karri, Jagadeshwari., College of Engineering and Computer Science, Department of Computer and Electrical Engineering and Computer Science
Abstract/Description: Cytogenetics is a study on the genetic considerations associated with structural and functional aspects of the cells with reference to chromosomal inclusions. Chromosomes are structures within the cells containing body's information in the form of strings of DNA. When atypical version or structural abnormality in one or more chromosomes prevails, it is defined as chromosomal aberrations (CA) depicting certain genetic pathogeny (known as genetic disorders). The present study assumes the...
Show moreCytogenetics is a study on the genetic considerations associated with structural and functional aspects of the cells with reference to chromosomal inclusions. Chromosomes are structures within the cells containing body's information in the form of strings of DNA. When atypical version or structural abnormality in one or more chromosomes prevails, it is defined as chromosomal aberrations (CA) depicting certain genetic pathogeny (known as genetic disorders). The present study assumes the presence of normal and abnormal chromosomal sets in varying proportions in the cytogenetic complex ; and, stochastical mixture theory is invoked to ascertain the information redundancy as a function of fractional abnormal chromosome population. This bioinformatic measure of redundancy is indicated as a track-parameter towards the progression of genetic disorder, for example, the growth of cancer. Lastly, using the results obtained, conclusions are enumerated, inferences are outlined and directions for future studies are considered.
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Date Issued: 2012
PURL: http://purl.flvc.org/FAU/3358597
Subject Headings: Medical genetics, Chromosome abnormalities, Cancer, Genetic aspects, Mutation (Biology), DNA damage
Format: Document (PDF)

Title: Developmental delays in methionine sulfoxide reductase mutants in Drosophila Melanogaster.
Creator: Hausman, William, Binninger, David, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Aging is a biological process that has many detrimental effects due to the accumulation of oxidative damage to key biomolecules due to the action of free radicals. Methionine sulfoxide reductase (Msr) functions to repair oxidative damage to methionine residues. Msr comes in two forms, MsrA and MsrB, each form has been shown to reduce a specific enantiomer of bound and free oxidized methionine. Effects of Msr have yet to be studied in the major developmental stages of Drosophila melanogaster...
Show moreAging is a biological process that has many detrimental effects due to the accumulation of oxidative damage to key biomolecules due to the action of free radicals. Methionine sulfoxide reductase (Msr) functions to repair oxidative damage to methionine residues. Msr comes in two forms, MsrA and MsrB, each form has been shown to reduce a specific enantiomer of bound and free oxidized methionine. Effects of Msr have yet to be studied in the major developmental stages of Drosophila melanogaster despite the enzymes elevated expression during these stages. A developmental timeline was determined for MsrA mutant, MsrB mutant, and double null mutants against a wild type control. Results show that the Msr double mutant is delayed approximately 20 hours in the early/mid third instar stage while each of the single mutants showed no significant difference to the wild type. Data suggests that the reasoning of this phenomenon is due to an issue gaining mass.
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Date Issued: 2014
PURL: http://purl.flvc.org/fau/fd/FA00004200, http://purl.flvc.org/fau/fd/FA00004200
Subject Headings: Aging -- Molecular aspects, Cellular signal transduction, Drosophila melanogaster -- Genetics, Mitochondrial pathology, Mutation (Biology), Oxidative stress
Format: Document (PDF)

Title: Identification and characterization of mutations in the Drosophila mitochondrial translation elongation factor iconoclast.
Creator: Trivigno, Catherine F., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Mitochondrial disorders resulting from defects in oxidative phosphorylation are the most common form of inherited metabolic disease. Mutations in the human mitochondrial translation elongation factor GFM1 have recently been shown to cause the lethal pediatric disorder Combined Oxidative Phosphorylation Deficiency Syndrome (COXPD1). Children harboring mutations in GFM1 exhibit severe developmental, metabolic and neurological abnormalities. This work describes the identification and extensive...
Show moreMitochondrial disorders resulting from defects in oxidative phosphorylation are the most common form of inherited metabolic disease. Mutations in the human mitochondrial translation elongation factor GFM1 have recently been shown to cause the lethal pediatric disorder Combined Oxidative Phosphorylation Deficiency Syndrome (COXPD1). Children harboring mutations in GFM1 exhibit severe developmental, metabolic and neurological abnormalities. This work describes the identification and extensive characterization of the first known mutations in iconoclast (ico), the Drosophila orthologue of GFM1. Expression of human GFM1 can rescue ico null mutants, demonstrating functional conservation between the human and fly proteins. While point mutations in ico result in developmental defects and death during embryogenesis, animals null for ico survive until the second or third instar larval stage. These results indicate that in addition to loss-of-function consequences, point mutations in ico appear to produce toxic proteins with antimorphic or neomorphic effects. Consistent with this hypothesis, transgenic expression of a mutant ICO protein is lethal when expressed during development and inhibits growth when expressed in wing discs. In addition, animals with a single copy of an ico point mutation are more sensitive to acute hyperthermic or hypoxic stress. Removal of the positively-charged tail of the protein abolishes the toxic effects of mutant ICO, demonstrating that this domain is necessary for the harmful gain-of-function phenotypes observed in ico point mutants., Further, expression of GFP-tagged constructs indicates that the C-terminal tail enhances ectopic nuclear localization of mutant ICO, suggesting that mislocalization of the protein may play a role in the antimorphic effects of mutant ICO. Taken together, these results illustrate that in addition to loss-of-function effects, gain-of-function effects can contribute significantly to the pathology caused by mutation in mitochondrial translation elongation factors.
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Date Issued: 2010
PURL: http://purl.flvc.org/FAU/2705081
Subject Headings: Drosophila melanogaster, Cytogenetics, Mutation (Biology), Mitochondrial DNA, Cell metabolism, Cellular signal transduction, Oxidation, Physiological, Genetic transcription, Regulation
Format: Document (PDF)

Title: Molecular and phenotypic characterization of MsrA MsrB mutants of Drosophila melanogaster.
Creator: Robbins, Kelli., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Aging is a multifactoral biological process of progressive and deleterious changes partially attributed to a build up of oxidatively damaged biomolecules resulting from attacks by free radicals. Methionine sulfoxide reductases (Msrs) are enzymes that repair oxidized methionine (Met) residues found in proteins. Oxidized Met produces two enantiomers, Met-S-(o) and Met-R-(o), reduced by MsrA and MsrB respectively. Unlike other model organisms, our MsrA null fly mutant did not display increased...
Show moreAging is a multifactoral biological process of progressive and deleterious changes partially attributed to a build up of oxidatively damaged biomolecules resulting from attacks by free radicals. Methionine sulfoxide reductases (Msrs) are enzymes that repair oxidized methionine (Met) residues found in proteins. Oxidized Met produces two enantiomers, Met-S-(o) and Met-R-(o), reduced by MsrA and MsrB respectively. Unlike other model organisms, our MsrA null fly mutant did not display increased sensitivity to oxidative stress or shortened lifespan, suggesting that in Drosophila, having either a functional copy of either Msr is sufficient. Here, two Msr mutant types were phenotypically assayed against isogenic controls. Results suggest that only the loss of both MsrA and MsrB produces increased sensitivity to oxidative stress and shortened lifespan, while locomotor defects became more severe with the full Msr knockout fly.
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Date Issued: 2009
PURL: http://purl.flvc.org/FAU/359920
Subject Headings: Genetic regulation, Oxidation-reduction reaction, Proteins, Chemical modification, Aging, Molecular aspects, Mutation (Biology), Cell metabolism, Mitochondrial DNA
Format: Document (PDF)

Title: Role of methionine sulfoxide reductase in thermal-induced spreading depression coma in Drosophila melanogaster.
Creator: Schey, Karin., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Drosophila melanogaster encounter periods of increased temperature or decreased oxygen in its native environment. One consequence of these environmental stresses is increased production of reactive oxygen species that damage major molecules within cells. Another consequence is that flies fall into a protective coma where biological functions are minimized to conserve energy expenditures. This biological phenomenon is called spreading depression. The overarching aim of this project is to...
Show moreDrosophila melanogaster encounter periods of increased temperature or decreased oxygen in its native environment. One consequence of these environmental stresses is increased production of reactive oxygen species that damage major molecules within cells. Another consequence is that flies fall into a protective coma where biological functions are minimized to conserve energy expenditures. This biological phenomenon is called spreading depression. The overarching aim of this project is to determine if methionine sulfoxide reductases affect entrance or exit from the protective coma induced by acute thermal stress. The data revealed that complete deficiency of Msr in young flies causes a faster induction of the coma. In both young and old flies, Msr does not affect average recovery time but does affect the pattern of recovery from coma. Entrance into the coma is age dependent with young flies maintaining activity longer than before entering into the coma as compared to old flies.
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Date Issued: 2012
PURL: http://purl.flvc.org/FAU/3355873
Subject Headings: Cellular signal transduction, Proteins, Chemical modification, Spreading cortical depression, Oxidation-reduction reaction, Aging, Molecular aspects, Mutation (Biology)
Format: Document (PDF)

Title: Methionine sulfoxide reductase (MSR) modulates lifespan andLocomotion in drosophila melanogaster.
Creator: Bruce, Lindsay, Binninger, David, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Oxidative stress is considered a major factor in the etiology of age related diseases and the aging process itself. Organisms have developed mechanisms to protect against oxidative damage resulting from increased production of reactive oxygen species during aging. One of the major antioxidant systems is the methionine sulfoxide reductase (Msr) enzyme family. The two major Msr enzymes, MsrA and MsrB, can stereospecifically reduce the S and R epimers, respectively, of methionine sulfoxide in...
Show moreOxidative stress is considered a major factor in the etiology of age related diseases and the aging process itself. Organisms have developed mechanisms to protect against oxidative damage resulting from increased production of reactive oxygen species during aging. One of the major antioxidant systems is the methionine sulfoxide reductase (Msr) enzyme family. The two major Msr enzymes, MsrA and MsrB, can stereospecifically reduce the S and R epimers, respectively, of methionine sulfoxide in proteins back to methionine. This study, using Drosophila melanogaster, decribes the first animal system lacking both MsrA and MsrB. The loss of either MsrA or MsrB had no effect on lifespan in Drosophila, but loss of MsrB results in a slight decrease in locomotor activity from middle age onward. Double mutants lacking both forms of Msr have a significantly decreased lifespan and decreased locomotor activity at all ages examined. The double Msr mutants had no detectable increase in protein oxidation or decrease in mitochondrial function and were not more sensitive to oxidative stress. These results suggested that other cellular antioxidant systems were protecting the flies against oxidative damage and the decreased life span observed in the double knockouts was not due to widespread oxidative damage. However, one cannot exclude limited oxidative damage to a specific locus or cell type. In this regard, it was observed that older animals, lacking both MsrA and MsrB, have significantly reduced levels of dopamine, suggesting there might be oxidative damage to the dopaminergic neurons. Preliminary results also suggest that the ratio of F to G actin is skewed towards G actin in all mutants. The present results could have relevance to the loss of dopaminergic neurons in Parkinson’s disease.
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Date Issued: 2015
PURL: http://purl.flvc.org/fau/fd/FA00004431, http://purl.flvc.org/fau/fd/FA00004431
Subject Headings: Aging -- Molecular aspects, Cellular signal transduction, Drosophila melanogaster -- Genetics, Mitochondrial pathology, Mutation (Biology), Oxidative stress, Proteins -- Chemical modification
Format: Document (PDF)

Title: Phenotypic and behavioral effects of methionine sulfoxide reductase deficiency and oxidative stress in Drosophila melanogaster.
Creator: Mulholland, Kori., Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: Harman's theory of aging proposes that a buildup of damaging reactive oxygen species (ROS) is one of the primary causes of the deleterious symptoms attributed to aging. Cellular defenses in the form of antioxidants have evolved to combat ROS and reverse damage; one such group is the methionine sulfoxide reductases (Msr), which function to reduce oxidized methionine. MsrA reduces the S enantiomer of methionine sulfoxide, Met-S-(o), while MsrB reduces the R enantiomer, Met-R-(o). The focus of...
Show moreHarman's theory of aging proposes that a buildup of damaging reactive oxygen species (ROS) is one of the primary causes of the deleterious symptoms attributed to aging. Cellular defenses in the form of antioxidants have evolved to combat ROS and reverse damage; one such group is the methionine sulfoxide reductases (Msr), which function to reduce oxidized methionine. MsrA reduces the S enantiomer of methionine sulfoxide, Met-S-(o), while MsrB reduces the R enantiomer, Met-R-(o). The focus of this study was to investigate how the absence of one or both forms of Msr affects locomotion in Drosophila using both traditional genetic mutants and more recently developed RNA interference (RNAi) strains. Results indicate that lack of MsrA does not affect locomotion. However, lack of MsrB drastically reduces rates of locomotion in all age classes. Furthermore, creation of an RNAi line capable of knocking down both MsrA and MsrB in progeny was completed.
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Date Issued: 2013
PURL: http://purl.flvc.org/fcla/dt/3362558
Subject Headings: Drosophila melanogaster, Genetics, Aging, Molecular aspects, Oxidative stress, Mitochondrial pathology, Cellular signal transduction, Oxidation-reduction reaction, Biochemical markers, Mutation (Biology)
Format: Document (PDF)

Title: Reduced Reproductivity and Larval Locomotion in the Absence of Methionine Sulfoxide Reductase in Drosophila.
Creator: Singkornrat, Diana, Binninger, David, Florida Atlantic University, Charles E. Schmidt College of Science, Department of Biological Sciences
Abstract/Description: The inevitable aging process can be partially attributed to the accumulation of oxidative damage that results from the action of free radicals. Methionine sulfoxide reductases (Msr) are a class of enzymes that repair oxidized methionine residues. The two known forms of Msr are MsrA and MsrB which reduce the R- and S- enantiomers of methionine sulfoxide, respectively. Our lab has created the first genetic animal model that is fully deficient for any Msr activity. Previously our lab showed that...
Show moreThe inevitable aging process can be partially attributed to the accumulation of oxidative damage that results from the action of free radicals. Methionine sulfoxide reductases (Msr) are a class of enzymes that repair oxidized methionine residues. The two known forms of Msr are MsrA and MsrB which reduce the R- and S- enantiomers of methionine sulfoxide, respectively. Our lab has created the first genetic animal model that is fully deficient for any Msr activity. Previously our lab showed that these animals exhibit a 20 hour delay in development of the third instar larvae (unpublished data). My studies have further shown that the prolonged third-instar stage is due to a reduced growth rate associated with slower food intake and a markedly slower motility. These Msr-deficient animals also exhibit decreased egg-laying that can be attributed to a lack of female receptivity to mating.
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Date Issued: 2016
PURL: http://purl.flvc.org/fau/fd/FA00004777, http://purl.flvc.org/fau/fd/FA00004777
Subject Headings: Proteins--Chemical modification., Oxidative stress., Oxidation-reduction reaction., Drosophila melanogaster--Genetics., Mitochondrial pathology., Cellular signal transduction., Mutation (Biology), Aging--Molecular aspects.
Format: Document (PDF)

FAU Theses and Dissertations (9)	+ -
Honors Student Theses (3)	+ -

Mutation (Biology) (12)	+ -
Aging (5)	+ -
Cell metabolism (5)	+ -
Cellular signal transduction (5)	+ -
Molecular aspects (5)	+ -

Mitochondrial DNA (3)	+ -
Mitochondrial pathology (3)	+ -
Oxidation-reduction reaction (3)	+ -
Oxidative stress (3)	+ -
Aging -- Molecular aspects (2)	+ -
Chemical modification (2)	+ -
Cytogenetics (2)	+ -
Drosophila melanogaster (2)	+ -
Medical genetics (2)	+ -
Microbial genetics (2)	+ -
Proteins (2)	+ -
Research (2)	+ -
Yeast fungi (2)	+ -
Aging--Molecular aspects. (1)	+ -
Biochemical markers (1)	+ -
Cancer (1)	+ -
Cell membranes (1)	+ -
Cellular signal transduction. (1)	+ -
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DNA damage (1)	+ -
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Drosophila melanogaster--Genetics. (1)	+ -
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thesis (3)	+ -

Charles E. Schmidt College of Science (7)	+ -
Department of Biological Sciences (7)	+ -
Florida Atlantic University (4)	+ -
Binninger, David (3)	+ -
Harriet L. Wilkes Honors College (3)	+ -

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