Matching Items (85)
Description
In eukaryotes, most messenger RNA precursors (pre-mRNA) undergo extensive processing, leading to the cleavage of the transcript followed by the addition of a poly(A) tail. This process is executed by a large complex known as the Cleavage and Polyadenylation Complex (CPC). Its central subcomplex, the Cleavage and Polyadenylation Specificity Factor (CPSF) complex is responsible for recognizing a short hexameric element AAUAAA located at the 3’end in the nascent mRNA molecule and catalyzing the pre-mRNA cleavage. In the round nematode C. elegans, the cleavage reaction is executed by a subunit of this complex named CPSF3, a highly conserved RNA endonuclease. While the crystal structure of its human ortholog CPSF73 has been recently identified, we still do not understand the molecular mechanisms and sequence specificity used by this protein to induce cleavage, which in turn would help to understand how this process is executed in detail. Additionally, we do not understand in additional factors are needed for this process. In order to address these issues, we performed a comparative analysis of the CPSF3 protein in higher eukaryotes to identify conserved functional domains. The overall percent identities for members of the CPSF complex range from 33.68% to 56.49%, suggesting that the human and C. elegans orthologs retain a high level of conservation. CPSF73 is the protein with the overall highest percent identity of the CPSF complex, with its active site-containing domain possessing 74.60% identity with CPSF3. Additionally, we gathered and expressed using a bacterial expression system CPSF3 and a mutant, which is unable to perform the cleavage reaction, and developed an in vitro cleavage assay to test whether CPSF3 activity is necessary and sufficient to induce nascent mRNA cleavage. This project establishes tools to better understand how CPSF3 functions within the CPC and sheds light on the biology surrounding the transcription process as a whole.
ContributorsGallante, Christina (Author) / Mangone, Marco (Thesis director) / Sharma, Shalini (Committee member) / Hrach, Heather (Committee member) / School of Life Sciences (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
The successful reduction of CO2 and protons by a light-induced cobalt porphyrin/cytb562 hybrid metalloenzyme in water is reported. Incorporation of the porphyrin into a protein scaffold results in increases in CO and H2 production over naked porphyrin. Rational point mutations to the CoPPIX binding site of cytb562 modulate production, indicating possible further improvements in catalytic activity.
ContributorsGwerder, Noah D (Author) / Ghirlanda, Giovanna (Thesis director) / Williams, Peter (Committee member) / Mangone, Marco (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
Duchenne muscular dystrophy (DMD) is a lethal, X-linked disease which occurs in approximately 1 in 3,500 male births. This disease is characterized by progressive muscle wasting and causes premature death. One of the earliest symptoms of this disease is mitochondrial dysfunction. Dystrophin is a protein found under the sarcolemma. The N terminus binds to actin and the C terminus binds to dystrophin glycoprotein complex (DGC). DMD is caused by mutations in the dystrophin gene. C. elegans possess an ortholog of dystrophin, DYS-1. Though there is evidence that C. elegans can be used as a model organism to model DMD, nematode DGC has not been well characterized. Additionally, while we know that mitochondrial dysfunction has been found in humans and other model organisms, this has not been well defined in C. elegans. In order to address these issues, we crossed the SJ4103 worm strain (myo-3p::GFP(mit)) with dys-1(cx18) in order to visualize and quantify changes in mitochondria in a dys-1 background. SJ4103;cx18 nematodes were found to have less mitochondrial than SJ4103 which suggests mitochondrial dysfunction does occur in dys-1 worms. Furthermore, mitochondrial dysfunction was studied by knocking down members of the DGC, dys-1, dyb-1, sgn-1, sgca-1, and sgcb-1 in SJ4103 strain. Knock down of each gene resulted in decrease in abundance of mitochondria which suggests that each member of the DGC contributes to the overall health of nematode muscle. The ORF of dyb-1 was successfully cloned and tagged with GFP in order to visualize this DGC member C. elegans. Imaging of the transgenic dyb-1::GFP worm shows green fluoresce expressed in which suggests that dyb-1 is a functional component of the muscle fibers. This project will enable us to better understand the effects of dystrophin deficiency on mitochondrial function as well as visualize the expression of certain members of the DGC in order to establish C. elegans as a good model organism to study this disease.
ContributorsObrien, Shannon Nishino (Author) / Mangone, Marco (Thesis director) / Newbern, Jason (Committee member) / Hrach, Heather (Committee member) / School of Molecular Sciences (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Cleavage and polyadenylation is a step in mRNA processing in which the 3’UTR is cleaved and a polyA tail is added to create a final mature transcript. This process relies on RNA sequence elements that guide a large multimeric protein complex named the Cleavage and Polyadenylation Complex to dock on the 3’UTR and execute the cleavage reaction. Interactions of the complex with the RNA and specific dynamics of complex recruitment and formation still remain largely uncharacterized. In our lab we have identified an Adenosine residue as the nucleotide most often present at the cleavage site, although it is unclear whether this specific element is a required instructor of cleavage and polyadenylation. To address whether the Adenosine residue is necessary and sufficient for the cleavage and polyadenylation reaction, we mutated this nucleotide at the cleavage site in three C. elegans protein coding genes, forcing the expression of these wt and mutant 3’UTRs, and studied how the cleavage and polyadenylation machinery process these genes in vivo. We found that interrupting the wt sequence elements found at the cleavage site interferes with the cleavage and polyadenylation reaction, suggesting that the sequence close to the end of the transcript plays a role in modulating the site of the RNA cleavage. This activity is also gene-specific. Genes such as ges-1 showed little disruption in the cleavage of the transcript, with similar location occurring in both the wt and mutant 3’UTRs. On the other hand, mutation of the cleavage site in genes such as Y106G6H.9 caused the activation of new cryptic cleavage sites within the transcript. Taken together, my experiments suggest that the sequence elements at the cleavage site somehow participate in the reaction to guide the cleavage reaction to occur at an exact site. This work will help to better understand the mechanisms of transcription termination in vivo and will push forward research aimed to study post-transcriptional gene regulation in eukaryotes.
ContributorsSteber, Hannah Suzanne (Author) / Mangone, Marco (Thesis director) / Harris, Robin (Committee member) / LaBaer, Joshua (Committee member) / School of Life Sciences (Contributor, Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Cultural Perceptions of Leisure and Well-being in Rock Climbing Communities of Peru and Arizona, USA
Description
In December of 2015, I made my way to rural Peru for a few weeks, my first visit to South America. While I was there, I observed a devotion to family and leisure activity, topics that were not heavily prioritized in my experience in Arizona. Upon my return, I became more involved in leisure activities, particularly running, hiking, yoga, and climbing. These involvements noticeably benefitted my health and well-being. The way the Peruvians I met prioritized these subjects fascinated me, and I wanted to study this difference between Arizona and Peru. In July of 2017, I returned to Peru for a semester abroad with my bags packed and the following research questions: 1) Are differences in motivation for rock climbing between Arizona and Peruvian climbers associated with cultural values? 2) Do leisure activities and the amount of time spent on them have an effect on quality of life? 3) Does the degree of climbing specialization impact perceptions of well-being? 4) What characteristics impact perceptions of quality of life among climbers? Are these characteristics affected by country of origin? My prediction was that Peruvians had higher quality of life due to their emphasis on leisure. Through this study, I learned that this conclusion was not as simple as I anticipated.
ContributorsMatta, Samantha Tania (Author) / Hultsman, Wendy (Thesis director) / Sampson, David (Committee member) / Lee, Rebecca (Committee member) / College of Integrative Sciences and Arts (Contributor) / School of Molecular Sciences (Contributor) / Division of Teacher Preparation (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
This is the study of Acute Impact of Ujjayi Yogic Pranayama vs Aerobic Exercise on Cognitive Performance and Short Term Memory. The purpose of this research was to compare two forms of exercise and their effects on someone's cognitive performance and short term memory. The research was performed in an acute setting were both exercises was conducted in under 15 minutes of active participation. The research question was; will aerobic exercise or the Pranayama breathing exercise provide better results and demonstrate a more effective way to increase the cognitive performance and short term memory for a college student aged 18-30. This was accomplished by using an aerobic exercise on an elliptical machine and then participating in the breathing exercise for 10 minutes in both scenarios. This study had two scenarios. Each scenario had a preliminary cognitive performance and short-term memory, post-Ujjayi exercise had a cognitive performance and short-term memory and a post-aerobic exercise had a cognitive performance and short-term memory. There was an hour break between Ujjayi exercise and aerobic exercise in both scenarios to prevent any type of bias. Scenario 1 had these three settings but the students were not given a breakfast supplement. In Scenario 2 the students were given a break supplement and followed the same procedures as scenario 1. There were 25 students for scenario 1 and 25 students for scenario 2. The students were allowed to participate in scenario 1 and 2 but it had to be a week after their first participation. All participants were originally signed up for scenario 1 and they could come back to perform scenario 2 a week later. The first scenario was completing the tests in the absence of food. Scenario two was completing the tests after having been given a Clif Bar to consume. The results of both of these scenarios showed that for cognitive performance and short term memory aerobic exercise had a beneficial impact on their performance. However, students who had a breakfast performed better on the preliminary tests and scored better after the yogic Ujjayi Pranayama exercise on their cognitive performance and short term memory tests. There was also a negligible difference between the test results after the preliminary tests and yogic Ujjayi Pranayama. However, in scenario one the overall tests scores for preliminary and yogic Ujjayi Pranayama were less than those in scenario two. Students who recorded that they were more actively engaging in regular physical exercise 3-7 days a week also did worse in scenario 1, but when presented with scenario 2 they scored equal with those who did not perform regular exercise. The overall purpose for this research was to find out how to increase cognitive performance and short term memory ability in college age students 18-30 in a short amount of time. The results of this study will be impactful for the future studies that will be focused on when comparing aerobic exercise and yogic pranayama.
ContributorsKopecky, Zachary (Co-author) / Enright, Roan (Co-author) / McILwraith, Heide (Thesis director) / Lee, Rebecca (Committee member) / College of Integrative Sciences and Arts (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
Description
This study confirms that there is stigma attached to how Somali-Americans perceive mental and emotional impairments compared to the perception of physical disabilities and impairments. More Somali-Americans are willing to seek help regarding their mental and physical health which is a positive step in improving the perceptions of Somali-Americans towards mental or emotional impairments and physical disabilities. Findings can contribute to the knowledge of health care professionals (i.e. nurses) in caring for patients identifying as Somali to promote culturally competent care.
ContributorsAden, Amina (Author) / Hosley, Brenda (Thesis director) / Lee, Rebecca (Committee member) / Lyles, Annmarie (Committee member) / Arizona State University. College of Nursing & Healthcare Innovation (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
Description
Nucleic acid polymers have numerous applications in both therapeutics and research to control gene expression and bind biologically relevant targets. However, due to poor biological stability their clinical applications are limited. Chemical modifications can improve both intracellular and extracellular stability and enhance resistance to nuclease degradation. To identify a potential candidate for a highly stable synthetic nucleic acid, the biostability of α-L-threofuranosyl nucleic acid (TNA) was evaluated under simulated biological conditions. TNA contains a four-carbon sugar and is linked by 2’, 3’ phosphodiester bonds. We hypothesized that this distinct chemical structure would yield greater nuclease resistance in human serum and human liver microsomes, which were selected as biologically relevant nuclease conditions. We found that TNA oligonucleotides remained undigested for 7 days in these conditions. In addition, TNA/DNA heteropolymers and TNA/RNA oligonucleotide duplexes displayed nuclease resistance, suggesting that TNA has a protective effect over DNA and RNA. In conclusion TNA demonstrates potential as a viable synthetic nucleic acid for use in numerous clinical and therapeutic applications.
ContributorsCulbertson, Michelle Catherine (Author) / Maley, Carlo (Thesis director) / Mangone, Marco (Committee member) / Larsen, Andrew (Committee member) / School of Molecular Sciences (Contributor) / School of International Letters and Cultures (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
Description
Research supports that music therapy can be used in multiple aspects of care for patients living within different environments. There is a gap in the literature when it comes to the impact of music sessions for older adults who do not have a diagnosed disease, therefore this study analyzes this population specifically. This study examines music therapy and its effects on anxiety and depression in adults aged 65 or older living in independent living homes. The adults participated in a mixed-methods study over the span of one month examining music as an intervention to decrease anxiety and depression. Each subject consented into the study, completed a demographic survey, answered open-ended questions regarding their experience with anxiety/sadness and ways to cope, as well as Profile of Moods Scale (POMS) during the first session. On the last week of the study, the participants were asked to fill out the same POMS scale to evaluate whether music influenced anxiety and depression. There was limited evidence found in this study to support the use of music therapy as an intervention to decrease anxiety and depression in adults over the age of 65.
ContributorsWolfus, Sarah Ilyssa (Author) / Lee, Rebecca (Thesis director) / Larkey, Linda (Committee member) / Arizona State University. College of Nursing & Healthcare Innovation (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
The Centers for Disease Control and Prevention in the United States announced that there has been roughly a 50% increase in the prevalence of food allergies among people between the years of 1997 - 2011. A food allergy can be described as a medical condition where being exposed to a certain food triggers a harmful immune response in the body, known as an allergic reaction. These reactions can range from mild to fatal, and they are caused mainly by the top 8 major food allergens: dairy, eggs, peanuts, tree nuts, wheat, soy, fish, and shellfish. Food allergies mainly plague children under the age of 3, as some of them will grow out of their allergy sensitivity over time, and most people develop their allergies at a young age, and not when they are older. The rise in prevalence is becoming a frightening problem around the world, and there are emerging theories that are attempting to ascribe a cause. There are three well-known hypotheses that will be discussed: the Hygiene Hypothesis, the Dual-Allergen Exposure Hypothesis, and the Vitamin-D Deficiency Hypothesis. Beyond that, this report proposes that a new hypothesis be studied, the Food Systems Hypothesis. This hypothesis theorizes that the cause of the rise of food allergies is actually caused by changes in the food itself and particularly the pesticides that are used to cultivate it.
ContributorsCromer, Kelly (Author) / Lee, Rebecca (Thesis director) / MacFadyen, Joshua (Committee member) / Sanford School of Social and Family Dynamics (Contributor) / Dean, W.P. Carey School of Business (Contributor) / Barrett, The Honors College (Contributor)
Created2018-12