Identifying statistical dependence in genomic sequences via mutual information estimates

Questions of understanding and quantifying the representation and amount of information in organisms have become a central part of biological research, as they potentially hold the key to fundamental advances. In this paper, we demonstrate the use of information-theoretic tools for the task of identifying segments of biomolecules (DNA or RNA) that are statistically correlated. We develop a precise and reliable methodology, based on the notion of mutual information, for finding and extracting statistical as well as structural dependencies. A simple threshold function is defined, and its use in quantifying the level of significance of dependencies between biological segments is explored. These tools are used in two specific applications. First, for the identification of correlations between different parts of the maize zmSRp32 gene. There, we find significant dependencies between the 5' untranslated region in zmSRp32 and its alternatively spliced exons. This observation may indicate the presence of as-yet unknown alternative splicing mechanisms or structural scaffolds. Second, using data from the FBI's Combined DNA Index System (CODIS), we demonstrate that our approach is particularly well suited for the problem of discovering short tandem repeats, an application of importance in genetic profiling.Comment: Preliminary version. Final version in EURASIP Journal on Bioinformatics and Systems Biology. See http://www.hindawi.com/journals/bsb

Spliced alignment and its application in Arabidopsis thaliana

This thesis describes the development and biological applications of GeneSeqer, which is a homology-based gene prediction program by means of spliced alignment. Additionally, a program named MyGV was written in JAVA as a browser to visualize the output of GeneSeqer. In order to test and demonstrate the performance, GeneSeqer was utilized to map 176,915 Arabidopsis EST sequences on the whole genome of Arabidopsis thaliana, which consists of five chromosomes, with about 117 million base pairs in total. All results were parsed and imported into a MySQL database. Information that was inferred from the Arabidopsis spliced alignment results may serve as valuable resource for a number of projects of special scientific interest, such as alternative splicing, non-canonical splice sites, mini-exons, etc. We also built AtGDB (Arabidopsis thaliana Genome DataBase, http://www.plantgdb.org/AtGDB/) to interactively browse EST spliced alignments and GenBank annotations for the Arabidopsis genome. Moreover, as one application of the Arabidopsis EST mapping data, U12-type introns were identified from the transcript-confirmed introns in the Arabidopsis genome, and the characteristics of these minor class introns were further explored

Basecalling for Traces Derived for Multiple Templates

Three methods for analyzing sequencing traces derived from sequencing reactions containing two DNA templates are presented. All rely on alignment to a segment of assembled genomic sequence containing the original template sequence. Spliced alignment algorithms are used so that traces derived from processed mRNA can be analyzed. The main application of these techniques is the elucidation of alternately spliced transcripts. Several experimental verification of one of the techniques is presented including testing on a set of 48 alternately spliced targets from the human genome and 47 negative controls

Large-scale methods in computational genomics

The explosive growth in biological sequence data coupled with the design and deployment of increasingly high throughput sequencing technologies has created a need for methods capable of processing large-scale sequence data in a time and cost effective manner. In this dissertation, we address this need through the development of faster algorithms, space-efficient methods, and high-performance parallel computing techniques for some key problems in computational genomics;The first problem addressed is the clustering of DNA sequences based on a measure of sequence similarity. Our clustering method: (i) guarantees linear space complexity, in contrast to the quadratic memory requirements of previously developed methods; (ii) identifies sequence pairs containing long maximal matches in the decreasing order of their maximal match lengths in run-time proportional to the sum of input and output sizes; (iii) provides heuristics to significantly reduce the number of pairs evaluated for checking sequence similarity without affecting quality; and (iv) has parallel strategies that provide linear speedup and a proportionate reduction in space per processor. Our approach has significantly enhanced the problem size reach while also drastically reducing the time to solution;The next problem we address is the de novo detection of genomic repeats called Long Terminal Repeat (LTR) retrotransposons. Our algorithm guarantees linear space complexity and produces high quality candidates for prediction in run-time proportional to the sum of input and output sizes. Validation of our approach on the yeast genome demonstrates both superior quality and performance results when compared to previously developed software;In a genome assembly project, fragments sequenced from a target genome are computationally assembled into numerous supersequences called contigs , which are then ordered and oriented into scaffolds . In this dissertation, we introduce a new problem called retroscaffolding for scaffolding contigs based on the knowledge of their LTR retrotransposon content. Through identification of sequencing gaps that span LTR retrotransposons, retroscaffolding provides a mechanism for prioritizing sequencing gaps for finishing purposes;While most of the problems addressed here have been studied previously, the main contribution in this dissertation is the development of methods that can scale to the largest available sequence collections

Identification and Functional Annotation of Alternatively Spliced Isoforms

Alternative splicing is a key mechanism for increasing the complexity of transcriptome and proteome in eukaryotic cells. A large portion of multi-exon genes in humans undergo alternative splicing, and this can have significant functional consequences as the proteins translated from alternatively spliced mRNA might have different amino acid sequences and structures. The study of alternative splicing events has been accelerated by the next-generation sequencing technology. However, reconstruction of transcripts from short-read RNA sequencing is not sufficiently accurate. Recent progress in single-molecule long-read sequencing has provided researchers alternative ways to help solve this problem. With the help of both short and long RNA sequencing technologies, tens of thousands of splice isoforms have been catalogued in humans and other species, but relatively few of the protein products of splice isoforms have been characterized functionally, structurally and biochemically. The scope of this dissertation includes using short and long RNA sequencing reads together for the purpose of transcript reconstruction, and using high-throughput RNA-sequencing data and gene ontology functional annotations on gene level to predict functions for alternatively spliced isoforms in mouse and human. In the first chapter, I give an introduction of alternative splicing and discuss the existing studies where next generation sequencing is used for transcript identification. Then, I define the isoform function prediction problem, and explain how it differs from better known gene function prediction problem. In the second chapter of this dissertation, I describe our study where the overall transcriptome of kidney is studied using both long reads from PacBio platform and RNA-seq short reads from Illumina platform. We used short reads to validate full-length transcripts found by long PacBio reads, and generated two high quality sets of transcript isoforms that are expressed in glomerular and tubulointerstitial compartments. In the third chapter, I describe our generic framework, where we implemented and evaluated several related algorithms for isoform function prediction for mouse isoforms. We tested these algorithms through both computational evaluation and experimental validation of the predicted ‘responsible’ isoform(s) and the predicted disparate functions of the isoforms of Cdkn2a and of Anxa6. Our algorithm is the first effort to predict and differentiate isoform functions through large-scale genomic data integration. In the fourth chapter, I present the extension of isoform function prediction study to the protein coding isoforms in human. We used a similar multiple instance learning (MIL)-based approach for predicting the function of protein coding splice variants in human. We evaluated our predictions using literature evidence of ADAM15, LMNA/C, and DMXL2 genes. And in the fifth and final chapter, I give a summary of previous chapters and outline the future directions for alternatively spliced isoform reconstruction and function prediction studies.PHDBioinformaticsUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttps://deepblue.lib.umich.edu/bitstream/2027.42/144017/1/ridvan_1.pd

Unsupervised and semi-supervised training methods for eukaryotic gene prediction

This thesis describes new gene finding methods for eukaryotic gene prediction. The current methods for deriving model parameters for gene prediction algorithms are based on curated or experimentally validated set of genes or gene elements. These training sets often require time and additional expert efforts especially for the species that are in the initial stages of genome sequencing. Unsupervised training allows determination of model parameters from anonymous genomic sequence with. The importance and the practical applicability of the unsupervised training is critical for ever growing rate of eukaryotic genome sequencing. Three distinct training procedures are developed for diverse group of eukaryotic species. GeneMark-ES is developed for species with strong donor and acceptor site signals such as Arabidopsis thaliana, Caenorhabditis elegans and Drosophila melanogaster. The second version of the algorithm, GeneMark-ES-2, introduces enhanced intron model to better describe the gene structure of fungal species with posses with relatively weak donor and acceptor splice sites and well conserved branch point signal. GeneMark-LE, semi-supervised training approach is designed for eukaryotic species with small number of introns. The results indicate that the developed unsupervised training methods perform well as compared to other training methods and as estimated from the set of genes supported by EST-to-genome alignments. Analysis of novel genomes reveals interesting biological findings and show that several candidates of under-annotated and over-annotated fungal species are present in the current set of annotated of fungal genomes.Ph.D.Committee Chair: Mark Borodovky; Committee Member: Jung H. Choi; Committee Member: King Jordan; Committee Member: Leonid Bunimovich; Committee Member: Yury Chernof

Computational modeling of gene structure in Arabidopsis thaliana

Computational gene identification by sequence inspection remains a challenging problem. For a typical Arabidopsis thaliana gene with five exons, at least one of the exons is expected to have at least one of its borders predicted incorrectly by ab initio gene finding programs. More detailed analysis for individual genomic loci can often resolve the uncertainty on the basis of EST evidence or similarity to potential protein homologues. Such methods are part of the routine annotation process. However, because the EST and protein databases are constantly growing, in many cases original annotation must be re-evaluated, extended, and corrected on the basis of the latest evidence. The Arabidopsis Genome Initiative is undertaking this task on the whole-genome scale via its participating genome centers. The current Arabidopsis genome annotation provides an excellent starting point for assessing the protein repertoire of a flowering plant. More accurate whole-genome annotation will require the combination of high-throughput and individual gene experimental approaches and computational methods. The purpose of this article is to discuss tools available to an individual researcher to evaluate gene structure prediction for a particular locus

Cross-species network and transcript transfer

Metabolic processes, signal transduction, gene regulation, as well as gene and protein expression are largely controlled by biological networks. High-throughput experiments allow the measurement of a wide range of cellular states and interactions. However, networks are often not known in detail for specific biological systems and conditions. Gene and protein annotations are often transferred from model organisms to the species of interest. Therefore, the question arises whether biological networks can be transferred between species or whether they are specific for individual contexts. In this thesis, the following aspects are investigated: (i) the conservation and (ii) the cross-species transfer of eukaryotic protein-interaction and gene regulatory (transcription factor- target) networks, as well as (iii) the conservation of alternatively spliced variants. In the simplest case, interactions can be transferred between species, based solely on the sequence similarity of the orthologous genes. However, such a transfer often results either in the transfer of only a few interactions (medium/high sequence similarity threshold) or in the transfer of many speculative interactions (low sequence similarity threshold). Thus, advanced network transfer approaches also consider the annotations of orthologous genes involved in the interaction transfer, as well as features derived from the network structure, in order to enable a reliable interaction transfer, even between phylogenetically very distant species. In this work, such an approach for the transfer of protein interactions is presented (COIN). COIN uses a sophisticated machine-learning model in order to label transferred interactions as either correctly transferred (conserved) or as incorrectly transferred (not conserved). The comparison and the cross-species transfer of regulatory networks is more difficult than the transfer of protein interaction networks, as a huge fraction of the known regulations is only described in the (not machine-readable) scientific literature. In addition, compared to protein interactions, only a few conserved regulations are known, and regulatory elements appear to be strongly context-specific. In this work, the cross-species analysis of regulatory interaction networks is enabled with software tools and databases for global (ConReg) and thousands of context-specific (CroCo) regulatory interactions that are derived and integrated from the scientific literature, binding site predictions and experimental data. Genes and their protein products are the main players in biological networks. However, to date, the aspect is neglected that a gene can encode different proteins. These alternative proteins can differ strongly from each other with respect to their molecular structure, function and their role in networks. The identification of conserved and species-specific splice variants and the integration of variants in network models will allow a more complete cross-species transfer and comparison of biological networks. With ISAR we support the cross-species transfer and comparison of alternative variants by introducing a gene-structure aware (i.e. exon-intron structure aware) multiple sequence alignment approach for variants from orthologous and paralogous genes. The methods presented here and the appropriate databases allow the cross-species transfer of biological networks, the comparison of thousands of context-specific networks, and the cross-species comparison of alternatively spliced variants. Thus, they can be used as a starting point for the understanding of regulatory and signaling mechanisms in many biological systems.In biologischen Systemen werden Stoffwechselprozesse, Signalübertragungen sowie die Regulation von Gen- und Proteinexpression maßgeblich durch biologische Netzwerke gesteuert. Hochdurchsatz-Experimente ermöglichen die Messung einer Vielzahl von zellulären Zuständen und Wechselwirkungen. Allerdings sind für die meisten Systeme und Kontexte biologische Netzwerke nach wie vor unbekannt. Gen- und Proteinannotationen werden häufig von Modellorganismen übernommen. Demnach stellt sich die Frage, ob auch biologische Netzwerke und damit die systemischen Eigenschaften ähnlich sind und übertragen werden können. In dieser Arbeit wird: (i) Die Konservierung und (ii) die artenübergreifende Übertragung von eukaryotischen Protein-Interaktions- und regulatorischen (Transkriptionsfaktor-Zielgen) Netzwerken, sowie (iii) die Konservierung von Spleißvarianten untersucht. Interaktionen können im einfachsten Fall nur auf Basis der Sequenzähnlichkeit zwischen orthologen Genen übertragen werden. Allerdings führt eine solche Übertragung oft dazu, dass nur sehr wenige Interaktionen übertragen werden können (hoher bis mittlerer Sequenzschwellwert) oder dass ein Großteil der übertragenden Interaktionen sehr spekulativ ist (niedriger Sequenzschwellwert). Verbesserte Methoden berücksichtigen deswegen zusätzlich noch die Annotationen der Orthologen, Eigenschaften der Interaktionspartner sowie die Netzwerkstruktur und können somit auch Interaktionen auf phylogenetisch weit entfernte Arten (zuverlässig) übertragen. In dieser Arbeit wird ein solcher Ansatz für die Übertragung von Protein-Interaktionen vorgestellt (COIN). COIN verwendet Verfahren des maschinellen Lernens, um Interaktionen als richtig (konserviert) oder als falsch übertragend (nicht konserviert) zu klassifizieren. Der Vergleich und die artenübergreifende Übertragung von regulatorischen Interaktionen ist im Vergleich zu Protein-Interaktionen schwieriger, da ein Großteil der bekannten Regulationen nur in der (nicht maschinenlesbaren) wissenschaftlichen Literatur beschrieben ist. Zudem sind im Vergleich zu Protein-Interaktionen nur wenige konservierte Regulationen bekannt und regulatorische Elemente scheinen stark kontextabhängig zu sein. In dieser Arbeit wird die artenübergreifende Analyse von regulatorischen Netzwerken mit Softwarewerkzeugen und Datenbanken für globale (ConReg) und kontextspezifische (CroCo) regulatorische Interaktionen ermöglicht. Regulationen wurden dafür aus Vorhersagen, experimentellen Daten und aus der wissenschaftlichen Literatur abgeleitet und integriert. Grundbaustein für viele biologische Netzwerke sind Gene und deren Proteinprodukte. Bisherige Netzwerkmodelle vernachlässigen allerdings meist den Aspekt, dass ein Gen verschiedene Proteine kodieren kann, die sich von der Funktion, der Proteinstruktur und der Rolle in Netzwerken stark voneinander unterscheiden können. Die Identifizierung von konservierten und artspezifischen Proteinprodukten und deren Integration in Netzwerkmodelle würde einen vollständigeren Übertrag und Vergleich von Netzwerken ermöglichen. In dieser Arbeit wird der artenübergreifende Vergleich von Proteinprodukten mit einem multiplen Sequenzalignmentverfahren für alternative Varianten von paralogen und orthologen Genen unterstützt, unter Berücksichtigung der bekannten Exon-Intron-Grenzen (ISAR). Die in dieser Arbeit vorgestellten Verfahren, Datenbanken und Softwarewerkzeuge ermöglichen die Übertragung von biologischen Netzwerken, den Vergleich von tausenden kontextspezifischen Netzwerken und den artenübergreifenden Vergleich von alternativen Varianten. Sie können damit die Ausgangsbasis für ein Verständnis von Kommunikations- und Regulationsmechanismen in vielen biologischen Systemen bilden

Computational analysis of noncoding RNAs

Noncoding RNAs have emerged as important key players in the cell. Understanding their surprisingly diverse range of functions is challenging for experimental and computational biology. Here, we review computational methods to analyze noncoding RNAs. The topics covered include basic and advanced techniques to predict RNA structures, annotation of noncoding RNAs in genomic data, mining RNA-seq data for novel transcripts and prediction of transcript structures, computational aspects of microRNAs, and database resources.Austrian Science Fund (Schrodinger Fellowship J2966-B12)German Research Foundation (grant WI 3628/1-1 to SW)National Institutes of Health (U.S.) (NIH award 1RC1CA147187