Elongation Factor EF-1α from Saccharomyces Cerevisiae Effects Translational Accuracy PDF Download
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Author: Mark Gordon Sandbaken
Publisher:
ISBN:
Category :
Languages : en
Pages : 322
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Author: Mark Gordon Sandbaken
Publisher:
ISBN:
Category :
Languages : en
Pages : 322
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Author: Felix Schirmaler
Publisher:
ISBN:
Category :
Languages : en
Pages : 238
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Author: Dustin Howard Hite
Publisher:
ISBN:
Category :
Languages : en
Pages :
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The central dogma of biology states that DNA, the genetic information, is transcribed into RNA, an information containing intermediate, which is then translated into proteins, actionable molecules which perform the majority of tasks required for life. To synthesize proteins, the cell employs a massive, macromolecular machine, the ribosome, and a myriad of protein factors to successfully translate an mRNA. My graduate studies have focused both on the ribosome and the protein translation factors that interact with the ribosome to facilitate translation initiation, elongation, and termination. First, utilizing recent advances in high throughput sequencing, we discovered that sequencing of ribosome protected fragments could illuminate in vivo dynamics of ribosome structural changes in Saccharomyces cerevisiae. We demonstrated that the ribosome protects two distinct sizes of fragments and assigned each fragment population to approximate stages of the translation elongation cycle where large structural rearrangements of the ribosome are known to occur. Once these assignments were made, we were able to model elongation speed and demonstrated that, contrary to previous reports, tRNA abundance and codon optimality were not the major determinants of elongation speed; surprisingly our data indicated that the polarity of the amino acid being decoded dictated elongation rates under these conditions, with polar amino acids acting to slow elongation rates. This study also implicated Dom34, a known NO GO decay factor, as a novel component of canonical translation termination and ribosome recycling. Second, we used another genome-wide assay of translation, "gradient encoding" microarray analysis, to interrogate the genome-wide effects of depleting five individual translation factors. Based on the current understanding of the molecular mechanisms of each translation factor, we hypothesized that the depletion of each factor would result in differential translation of mRNAs based on the physical properties of each mRNA species. However, we were startled to observe that the translational program of S. cerevisiae was relatively unperturbed by the depletion of three initiation factors, one elongation factor, and one termination factor. Further investigation revealed that yeast were actively compensating for the deficiency of each factor by either increasing or decreasing translation initiation rates such that the depleted factor was no longer limiting. This tuning was mediated by changes in eIF2[alpha] phosphorylation levels, a known modulator of translation initiation. Overall, we have leveraged high throughput technologies to provide novel understanding of in vivo structural dynamics of the ribosome and reveal a novel, unexpected robustness of the translational program in S. cerevisiae.
Author: Chi-Ting Huang
Publisher:
ISBN:
Category : Saccharomyces cerevisiae
Languages : en
Pages : 244
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Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 998
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Author: Wanfu Hou
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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While we have learned that mRNA sequence strongly influences translation and co-translational pathways, the exact mechanisms by which this sequence and the RNA structures encoded impact these steps of gene expression are less understood. Therefore, more investigations are needed to reveal how translation elongation and efficiency is influenced by RNA sequence and how the affected translation regulates and determines a variety of co-translational pathways. In this dissertation, I explore the effects of translational kinetics on a series of co-translational pathways using the budding yeast Saccharomyces cerevisiae as a model organism. In Chapter 2, I developed an in-vivo elongation reporter in Saccharomyces cerevisiae to quantitatively monitor translation elongation duration and protein expression. Using this elongation reporter, I investigated the effects of elongation stalls induced by different types of genetic factors on gene expression and demonstrated that distinct ribosomal stalls may trigger distinct co-translational pathways. In Chapter 3, I studied co-translational mRNA localization to mitochondrion, and proposed that translational kinetics, such as ribosomal stall caused by polyprolines, play an important role in mediating co-translational import. In addition, I further studied the effects of elongation stalls on mitochondrial import stress and triggering of relevant quality control pathways. In Chapter 4, I investigated the mechanism of cytosolic mRNP granule formation under glucose deprivation condition. In this study, I use CRISPRi to knockdown expression of RVB2 and confirm its essential role in deciding the fate of mRNA localization and translatability after glucose depletion. Finally in Chapter 5, I address the enhancements made to the developed method, outline potential future directions for the research presented in this dissertation, and conclude with my final remarks.
Author:
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Category : Dissertations, Academic
Languages : en
Pages : 750
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Author: David C. Amberg
Publisher: CSHL Press
ISBN: 0879697288
Category : Genetics
Languages : en
Pages : 250
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"Methods in Yeast Genetics" is a course that has been offered annually at Cold Spring Harbor for the last 30 years. This provides a set of teaching experiments along with the protocols and recipes for the standard techniques and reagents used in the study of yeast biology.
Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1902
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Vols. for 1963- include as pt. 2 of the Jan. issue: Medical subject headings.
Author: Pedro A. Ortiz
Publisher:
ISBN:
Category :
Languages : en
Pages : 268
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