Browsing by Subject "secondary structure"
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Item Advances in diapriid (Hymenoptera: diapriidae) systematics, with contributions to cybertaxonomy and the analysis of rRNA sequence data(2009-05-15) Yoder, Matthew JonDiapriids (Hymenoptera: Diapriidae) are small parasitic wasps. Though found throughout the world they are relatively unknown. A framework for advancing diapriid systematics is developed by introducing a new web-based application/database capable of storing a broad range of systematic data, and the first molecular phylogeny specifically focused at examining intrafamilial relationships. In addition to these efforts, a description of a new taxon is provided. Several advantages of digital description, including linking descriptions to an ontology of morphological terms, are highlighted. The functionality of the database is further illustrated in the production of a catalog of diapriid host associations. The hosts database currently holds over 450 association records, for over 500 named taxa (parasitoids and hosts), and over 180 references. Diapriids are found to be primarily endoparasitoids of Diptera emerging from the host pupa. Phylogenetic inference for a molecular dataset of 28S and 18S rRNA sequence data, derived from a diverse selection of diapriids, is accomplished with a new suite of tools developed for handling complex rRNA datasets. Several parsimony-based methodologies, including an alignment-free method of analyzing multiple sequences, are reviewed and applied using the new software tools. Diapriid phylogenetic relationships are shown to be broadly congruent with existing morphology-based classifications. Methods for analyzing typically excluded sequence data are shown to recover phylogenetic signal that would otherwise be lost and the alignment-free method performed remarkably well in this regard. Empirically, phylogenetic approaches that incorporate structural data were not notably different than those that did not.Item Investigation of Peptide Folding by Nuclear Magnetic Resonance Spectroscopy(2012-07-16) Hwang, SoYounUnderstanding structure and folding of a protein is the key to understanding its biological function and potential role in diseases. Despite the importance of protein folding, a molecular level understanding of this process is still lacking. Solution-state nuclear magnetic resonance (NMR) is a powerful technique to investigate protein structure, dynamics, and folding mechanisms, since it provides residue-specific information. One of the major contributions that govern protein structure appears to be the interaction with the solvent. The importance of these interactions is particularly apparent in membrane proteins, which exist in an amphiphilic environment. Here, individual peptide fragments taken from the disulfide bond forming protein B (DsbB) were investigated in various solvents. The alpha-helical structures that were obtained, suggest that DsbB follows the two-stage model for folding. However, side chains of polar residues showed different conformations compared to the X-ray structure of fulllength protein, implying that polar side-chains may re-orient upon helix packing in order to form the necessary tertiary interactions that stabilize the global fold of DsbB. Model peptides in general represent attractive systems for the investigation of non-covalent interactions important for protein folding, including those with the solvent. NMR structures of the water soluble peptide, BBA5, were obtained in the presence an organic co-solvent, methanol. These structures indicate that the addition of methanol stabilizes an alpha-helix segment, but disrupts a hydrophobic cluster forming a beta-hairpin. Since dynamic effects reduce the ability for experimental observation of individual, bound solvent molecules, results were compared with molecular dynamics simulations. This comparison indicates that the observed effects of NMR structures are due to preferred binding of methanol and reduction of peptide-water hydrogen bonding. NMR structures, such as those determined here, represent a distribution of conformations under equilibrium. The dynamic process of protein unfolding can nevertheless be accessed through denaturation. A method was developed to probe thermal denaturation by measuring the temperature dependence of NOE intensity. Applied to a model peptide, trpzip4, it was confirmed that the beta-hairpin structure of this peptide is stabilized by the hydrophobic cluster formed by tryptophan residues. Together, the peptides investigated here illustrate the important roles that solvent-peptide interactions and side chain-side chain hydrophobic interactions play in forming stable secondary and tertiary structures.Item Structure-based methods for the phylogenetic analysis of ribosomal RNA molecules(Texas A&M University, 2005-11-01) Gillespie, Joseph JamesRibosomal RNA (rRNA) molecules form highly conserved secondary and tertiary structures via rRNA-rRNA and rRNA-protein interactions that collectively comprise the macromolecule that is the ribosome. Because of their cellular universality, rRNA molecules are commonly used for phylogeny estimations spanning all divergences of life. In this dissertation, I elucidate the structure of several rRNAs by analyzing multiply aligned sequences for basepair covariation and conserved higher order structural motifs. Specifically, I predict novel structures for expansion segments D2 and D3 of the nuclear large subunit rRNA (28S) and variable regions V4-V9 of the nuclear small subunit rRNA (18S) from from 249 galerucine leaf beetles (Coleoptera: Chrysomelidae). I describe a novel means for characterizing regions of alignment ambiguity that improves methods for retaining phylogenetic information without violating nucleotide positional homology. In the program PHASE, I explore a variety of RNA maximum likelihood models using the 28S rRNA dataset and discuss the utitilty of these models in light of their performance under Bayesian analysis. I conclude that seven-state models are likely the best models to use for phylogenetic estimation, although I cannot determine with confidence which of the two seven-state models (7A or 7D) is better. Evaluation of the unpaired sites within both rRNAs in Modeltest provided a similar model of evolution for these non-pairing regions (TrN+ I+G). In addition, a sequenced region of the mitochondrial cytochrome oxidase I gene (COI) from the galerucines was evaluated in Modeltest, with each codon position modeled separately (GTR+I+G for positions 1 and 2, GTR+G for position 3). The combined galerucine dataset (28S+18S rRNA helices, 28S+18S rRNA unpaired sites, COI 1st, 2nd and 3rd positions) provided for two mixedmodel Bayesian analysis of five discretely-modeled partitions (using 7A and 7D). The results of these analyses are compared with those obtained from equally weighted parsimony to provide a robust phylogenetic estimate of the Galerucinae and related leaf beetle taxa. Finally, the odd characteristics of strepsipteran 18S rRNA are evaluated through comparison of 12 strepsipterans with 163 structurally-aligned arthropod sequences. Among other interesting results, I identify errors in previously published strepsipteran sequences and predict structures not previously known from metazoan rRNA.