References of "May, Patrick 50002348"
Showing results 141 to 160 of 176
A Protein Prioritization Approach Tailored for the FA/BRCA Pathway
Haitjema, Anneke; Brandt, Bernd W.; Ameziane, Najim et al
in PLoS ONE (2013), 8(4), 62017
<sec><title/><p>Fanconi anemia (FA) is a heterogeneous recessive disorder associated with a markedly elevated risk to develop cancer. To date sixteen FA genes have been identified, three of which ... [more ▼]
<sec><title/><p>Fanconi anemia (FA) is a heterogeneous recessive disorder associated with a markedly elevated risk to develop cancer. To date sixteen FA genes have been identified, three of which predispose heterozygous mutation carriers to breast cancer. The FA proteins work together in a genome maintenance pathway, the so-called FA/BRCA pathway which is important during the <italic>S</italic> phase of the cell cycle. Since not all FA patients can be linked to (one of) the sixteen known complementation groups, new FA genes remain to be identified. In addition the complex FA network remains to be further unravelled. One of the FA genes, <italic>FANCI</italic>, has been identified via a combination of bioinformatic techniques exploiting FA protein properties and genetic linkage. The aim of this study was to develop a prioritization approach for proteins of the entire human proteome that potentially interact with the FA/BRCA pathway or are novel candidate FA genes. To this end, we combined the original bioinformatics approach based on the properties of the first thirteen FA proteins identified with publicly available tools for protein-protein interactions, literature mining (Nermal) and a protein function prediction tool (FuncNet). Importantly, the three newest FA proteins FANCO/RAD51C, FANCP/SLX4, and XRCC2 displayed scores in the range of the already known FA proteins. Likewise, a prime candidate FA gene based on next generation sequencing and having a very low score was subsequently disproven by functional studies for the FA phenotype. Furthermore, the approach strongly enriches for GO terms such as DNA repair, response to DNA damage stimulus, and cell cycle-regulated genes. Additionally, overlaying the top 150 with a haploinsufficiency probability score, renders the approach more tailored for identifying breast cancer related genes. This approach may be useful for prioritization of putative novel FA or breast cancer genes from next generation sequencing efforts.</p></sec> [less ▲]
Differentiated SH-SY5Y Cells as PD Model for Mitochondrial Dysfunction: From Whole Genome Sequencing to an Educated Design of High-Throughput Experiments
Antony, Paul ; Krishna, Abhimanyu ; May, Patrick et al
Poster (2013)
Objectives: Mitochondrial dysfunction is a cellular hallmark of Parkinson's disease (PD) and energetic stress of dopaminergic neurons appears to be a physiological risk factor for mitochondrial ... [more ▼]
Objectives: Mitochondrial dysfunction is a cellular hallmark of Parkinson's disease (PD) and energetic stress of dopaminergic neurons appears to be a physiological risk factor for mitochondrial dysfunction. It is however challenging to assess the high variety of factors regulating mitochondrial physiology in living neurons in a high throughput manner. To overcome this bottleneck, we established an analysis platform, using the neuroblastoma cell line SH-SY5Y. For the first time ever we have characterized the SH-SY5Y cell line in an integrated whole genome, transcriptome, and proteome approach. In addition, we show that neuronal differentiation improves the physiological properties of this experimental model for studying mitochondrial dysfunction in PD. Methods: Whole genome sequencing, RNA-Seq, qRT-PCR, MS, FRET using Voltage sensing proteins, Immunofluorescence, cytometry, and live cell imaging. Results: The integrated molecular characterization of SH-SY5Y uncovers the level of molecular network integrity and hence the relevance of this cell line for targeted studies in selected molecular processes. Furthermore, we dissect changes in mitochondrial and energetic stress factors during the process of neuronal differentiation. Conclusions: In terms of both morphology and energetic stress response, differentiated SH-SY5Y cells are more similar to dopaminergic neurons than their undifferentiated precursors. Thanks to dividing progenitors and the short duration of differentiation, combined with the use of specific endpoints analysed with high-content microscopy, our platform paves the route for high throughput experiments on a neuronal cell culture model for PD. Our genomic characterization and expression profiling of SH-SY5Y cells furthermore helps guiding the experimental design and interpretation of such studies. [less ▲]
A model microbial community for Eco-Systems Biology
Muller, Emilie ; Roume, Hugo ; Buschart, Anna et al
Objective Microbial communities (MCs) play crucial roles in human health and disease. In-depth characterization of the vast organismal and functional diversity of MCs is now facilitated by high-resolution ... [more ▼]
Objective Microbial communities (MCs) play crucial roles in human health and disease. In-depth characterization of the vast organismal and functional diversity of MCs is now facilitated by high-resolution molecular approaches. Systematic measurements are key for meaningful data integration, analysis and modeling. Based on a model MC from a biological wastewater treatment plant, we have developed a new framework based on wet- and dry-lab methods for the integrated analyses of MCs at the population- as well as at the community-level. Methods The overall methodological framework first relies on a standardised wet-lab procedure for the isolation of concomitant biomolecules, i.e., DNA, RNA, proteins and metabolites, from single undivided samples. Purified biomolecular fractions then are subjected to high-resolution omic analyses including metagenomics, metatranscriptomics, metaproteomics and (meta-) metabolomics. The resulting data form the input for integrated bioinformatic analyses. Population-level integrated omic analyses rely on a newly developed binning and re-assembly method, which yields near-complete genome reconstructions for dominant populations. Community-level analyses involve the reconstruction of community-wide metabolic networks. Functional omic data is then mapped onto these reconstructions and contextualized. Results Application of the population-centric workflow has allowed us to reconstruct and identify 10 major populations within the model MC and has led to the identification of a key generalist population, Candidatus Microthrix spp., within the community. Analysis of the community-wide metabolic networks has allowed the identification of keystone genes involved in lipid and nitrogen metabolism within the MC. Conclusions Our new methodological framework offers exciting new prospects for elucidating the functional relevance of specific populations and genes within MCs. The established workflows are now being applied to samples of biomedical research interest such as human gastrointestinal tract-derived samples. [less ▲]
Condensing the omics fog of microbial communities
Muller, Emilie ; Glaab, Enrico ; May, Patrick et al
in Trends in Microbiology (2013), 21(7), 325333
Natural microbial communities are ubiquitous, complex, heterogeneous and dynamic. Here, we argue that the future standard for their study will require systematic omic measurements of spatially and ... [more ▼]
Natural microbial communities are ubiquitous, complex, heterogeneous and dynamic. Here, we argue that the future standard for their study will require systematic omic measurements of spatially and temporally resolved unique samples in line with a discovery-driven planning approach. Resulting datasets will allow the generation of solid hypotheses about causal relationships and, thereby, will facilitate the discovery of previously unknown traits of specific microbial community members. However, to achieve this, solid wet-lab, bioinformatic and statistical methodologies are required to have the promises of the emerging field of Eco-Systems Biology come to fruition. [less ▲]
New insights into Chlamydomonas reinhardtii hydrogen production processes by combined microarray/RNA-seq transcriptomics.
Toepel, Jorg; Illmer-Kephalides, Maike; Jaenicke, Sebastian et al
in Plant Biotechnology Journal (2013)
Hydrogen production with Chlamydomonas reinhardtii induced by sulphur starvation is a multiphase process while the cell internal metabolism is completely remodelled. The first cellular response is ... [more ▼]
Hydrogen production with Chlamydomonas reinhardtii induced by sulphur starvation is a multiphase process while the cell internal metabolism is completely remodelled. The first cellular response is characterized by induction of genes with regulatory functions, followed by a total remodelling of the metabolism to provide reduction equivalents for cellular processes. We were able to characterize all major processes that provide energy and reduction equivalents during hydrogen production. Furthermore, C. reinhardtii showed a strong transcript increase for gene models responsible for stress response and detoxification of oxygen radicals. Finally, we were able to determine potential bottlenecks and target genes for manipulation to increase hydrogen production or to prolong the hydrogen production phase. The investigation of transcriptomic changes during the time course of hydrogen production in C. reinhardtii with microarrays and RNA-seq revealed new insights into the regulation and remodelling of the cell internal metabolism. Both methods showed a good correlation. The microarray platform can be used as a reliable standard tool for routine gene expression analysis. RNA-seq additionally allowed a detailed time-dependent study of gene expression and determination of new genes involved in the hydrogen production process. [less ▲]
Integrated omics highlights keystone genes in community-wide metabolic networks
Roume, Hugo ; Satagopam, Venkata ; Muller, Emilie et al
Systematic molecular measurements reveal key microbial populations driving community-wide phenotype
Muller, Emilie ; Pinel, Nicolás; May, Patrick et al
Natural microbial communities are heterogeneous and dynamic. Therefore, a major consideration for multiple omic data studies is the sample-to-sample heterogeneity, which can lead to inconsistent results ... [more ▼]
Natural microbial communities are heterogeneous and dynamic. Therefore, a major consideration for multiple omic data studies is the sample-to-sample heterogeneity, which can lead to inconsistent results if the different biomolecular fractions are obtained from distinct sub-samples. Conversely, systematic omic measurements, i.e. the standardised, reproducible and simultaneous measurement of multiple features from a single undivided sample, result in fully integrable datasets. Objective In order to prove the feasibility and benefits of such systematic measurements in the study of the respective contributions of different populations to the community-wide phenotype, we purified and analysed all biomolecular fractions, i.e. DNA, RNA, proteins and metabolites, obtained from a unique undivided sample of lipid accumulating microbial community (LAMC) from wastewater treatment plant and integrate the resulting datasets. Methods One time point of particular interest was first selected out of 4 LAMC samples for its high diversity and strong lipid accumulation phenotype. Then, the systematic measurement strategy was applied to the selected undivided LAMC sample and the purified biomolecules were analysed by high-throughput techniques. DNA and RNA sequencing reads were assembled at the population-level using different binning strategies. A database, containing predicted proteins, was constructed to identify the detected peptides. Finally, all biomolecular information was mapped onto the assembled composite genomes to identify the precise roles of the different populations in the community-wide lipid accumulation phenotype. Results Metabolomics and 16S diversity analyses were used to select the sample of highest interest for detailed analysis. The systematic measurements of the selected sample followed by data integration have allowed us to probe the functional relevance of the population-level composite genomes, leading to the identification of the LAMC key players. Conclusion As community phenotype is not the sum of the different partner phenotypes, understanding a microbial community system requires more than the study of isolated organisms. Even if both approaches are complementary, top-down systematic approached only provides a holistic perspective of micro-ecological processes. [less ▲]
Linking mixed microbial community phenotype to individual genotypes
Biological wastewater treatment is arguably the most widely used biotechnological process on Earth. Wastewater also represents a valuable energy commodity that is currently not being harnessed ... [more ▼]
Biological wastewater treatment is arguably the most widely used biotechnological process on Earth. Wastewater also represents a valuable energy commodity that is currently not being harnessed comprehensively. Mixed microbial communities that naturally occur at the air-water interface of certain biological wastewater treatment systems accumulate excess long chain fatty acids intracellularly. This phenotypic trait may potentially be exploited for the transformation of lipid-rich wastewater into biodiesel (fatty acid methyl esters). Using a molecular Eco-Systems Biology approach, we are studying which genes contribute to the lipid accumulation phenotype and, thus, overall community function. We first compared the lipid accumulation phenotype to the structure of lipid accumulating communities from a local wastewater treatment plant by coupled deep sequencing of the 16S rRNA locus, metagenome sequencing and metabolomic analysis of 4 biological replicates sampled at 4 different time points. Based on the results of these analyses and in order to obtain a detailed view of the structure and function of the respective microbial communities, metagenomic, metatranscriptomic, metaproteomic and (meta-)metabolomic analyses were completed for a single representative biological sample of highest interest. In order to facilitate meaningful data integration of this highly heterogeneous consortium, biomolecular fractions used for the omic analyses were extracted from a unique single sample using a recently developed biomolecular isolation protocol. The coupled survey and the comparative metagenomic analysis demonstrate that the communities change significantly from dates with warm water temperatures to cold water temperatures while alpha diversity remains stable. In the winter period, this switch results in a strong enrichment of a bacterial genus well known to accumulate intracellular lipids, namely Microthrix spp., a representative genome of which has recently been sequenced by us. Correlation networks based on microorganism and concomitant intra- and extra-cellular metabolite abundances provides an overview of organisms potentially involved in the community-wide lipid accumulating phenotype. A sample with the highest abundance of Microthrix spp. was subsequently chosen for the construction of a community-wide metabolic model using metagenomic, metatranscriptomic, metaproteomic and (meta-)metabolomics data. Based on these omic datasets, expressed enzyme variants linked to the lipid accumulation phenotype have been identified and are currently undergoing in vitro characterization. Meta-omic analyses offer exciting prospects for elucidating the genetic blueprints and the functional relevance of specific populations within microbial communities. Consequently, connecting the overall community phenotype to specific genotypes will allow much needed fundamental ecological understanding of microbial community and population dynamics, particularly in relation to environment-driven demography changes leading to tipping points and catastrophic bifurcations. [less ▲]
Give It AGO: The Search for miRNA-Argonaute Sorting Signals in Arabidopsis thaliana Indicates a Relevance of Sequence Positions Other than the 5'-Position Alone.
Thieme, Christoph J.; Schudoma, Christian; May, Patrick et al
in Frontiers in Plant Science (2012), 3
The specific recognition of miRNAs by Argonaute (AGO) proteins, the effector proteins of the RNA-induced silencing complex, constitutes the final step of the biogenesis of miRNAs and is crucial for their ... [more ▼]
The specific recognition of miRNAs by Argonaute (AGO) proteins, the effector proteins of the RNA-induced silencing complex, constitutes the final step of the biogenesis of miRNAs and is crucial for their target interaction. In the genome of Arabidopsis thaliana (Ath), 10 different AGO proteins are encoded and the sorting decision, which miRNA associates with which AGO protein, was reported to depend exclusively on the identity of the 5'-sequence position of mature miRNAs. Hence, with only four different bases possible, a 5'-position-only sorting signal would not suffice to specifically target all 10 different AGOs individually or would suggest redundant AGO action. Alternatively, other and as of yet unidentified sorting signals may exist. We analyzed a dataset comprising 117 Ath-miRNAs with clear sorting preference to either AGO1, AGO2, or AGO5 as identified in co-immunoprecipitation experiments combined with sequencing. While mutual information analysis did not identify any other single position but the 5'-nucleotide to be informative for the sorting at sufficient statistical significance, significantly better than random classification results using Random Forests nonetheless suggest that additional positions and combinations thereof also carry information with regard to the AGO sorting. Positions 2, 6, 9, and 13 appear to be of particular importance. Furthermore, uracil bases at defined positions appear to be important for the sorting to AGO2 and AGO5, in particular. No predictive value was associated with miRNA length or base pair binding pattern in the miRNA:miRNA* duplex. From inspecting available AGO gene expression data in Arabidopsis, we conclude that the temporal and spatial expression profile may also contribute to the fine-tuning of miRNA sorting and function. [less ▲]
Computation and Visualization of Protein Topology Graphs Including Ligand Information
Schäfer, Tim; May, Patrick ; Koch, Ina
in Böcker, Sebasttian; Hufsky, Franziska; Scheubert, Kerstin (Eds.) et al German Conference on Bioinformatics 2012 (2012)
Integration of proteomic and metabolomic profiling as well as metabolic modeling for the functional analysis of metabolic networks.
May, Patrick ; Christian, Nils ; Ebenhoh, Oliver et al
in Methods in Molecular Biology (2011), 694
The integrated analysis of different omics-level data sets is most naturally performed in the context of common process or pathway association. In this chapter, the two basic approaches for a metabolic ... [more ▼]
The integrated analysis of different omics-level data sets is most naturally performed in the context of common process or pathway association. In this chapter, the two basic approaches for a metabolic pathway-centric integration of proteomics and metabolomics data are described: the knowledge-based approach relying on existing metabolic pathway information, and a data-driven approach that aims to deduce functional (pathway) associations directly from the data. Relevant algorithmic approaches for the generation of metabolic networks of model organisms, their functional analysis, database resources, visualization and analysis tools will be described. The use of proteomics data in the process of metabolic network reconstruction will be discussed. [less ▲]
Comparative analysis of miRNAs and their targets across four plant species.
Lenz, Dorina; May, Patrick ; Walther, Dirk
in BMC Research Notes (2011), 4
BACKGROUND: MicroRNA (miRNA) mediated regulation of gene expression has been recognized as a major posttranscriptional regulatory mechanism also in plants. We performed a comparative analysis of miRNAs ... [more ▼]
BACKGROUND: MicroRNA (miRNA) mediated regulation of gene expression has been recognized as a major posttranscriptional regulatory mechanism also in plants. We performed a comparative analysis of miRNAs and their respective gene targets across four plant species: Arabidopsis thaliana (Ath), Medicago truncatula(Mtr), Brassica napus (Bna), and Chlamydomonas reinhardtii (Cre). RESULTS: miRNAs were obtained from mirBase with 218 miRNAs for Ath, 375 for Mtr, 46 for Bna, and 73 for Cre, annotated for each species respectively. miRNA targets were obtained from available database annotations, bioinformatic predictions using RNAhybrid as well as predicted from an analysis of mRNA degradation products (degradome sequencing) aimed at identifying miRNA cleavage products. On average, and considering both experimental and bioinformatic predictions together, every miRNA was associated with about 46 unique gene transcripts with considerably variation across species. We observed a positive and linear correlation between the number miRNAs and the total number of transcripts across different plant species suggesting that the repertoire of miRNAs correlates with the size of the transcriptome of an organism. Conserved miRNA-target pairs were found to be associated with developmental processes and transcriptional regulation, while species-specific (in particular, Ath) pairs are involved in signal transduction and response to stress processes. Conserved miRNAs have more targets and higher expression values than non-conserved miRNAs. We found evidence for a conservation of not only the sequence of miRNAs, but their expression levels as well. CONCLUSIONS: Our results support the notion of a high birth and death rate of miRNAs and that miRNAs serve many species specific functions, while conserved miRNA are related mainly to developmental processes and transcriptional regulation with conservation operating at both the sequence and expression level. [less ▲]
Distribution pattern of small RNA and degradome reads provides information on miRNA gene structure and regulation.
Branscheid, Anja; Devers, Emanuel A.; May, Patrick et al
in Plant signaling & behavior (2011), 6(10), 1609-11
Plant microRNAs (miRNAs) have an impact in the regulation of several biological processes such as development, growth and metabolism by negatively controlling gene expression at the post-transcriptional ... [more ▼]
Plant microRNAs (miRNAs) have an impact in the regulation of several biological processes such as development, growth and metabolism by negatively controlling gene expression at the post-transcriptional level. However, the role of these small molecules in the symbiotic interaction of plant roots and arbuscular mycorrhizal (AM) fungi remained elusive. To elucidate the role of miRNAs during AM symbiosis we used a deep sequencing approach to analyze the small RNA and degradome sequence tags of Medicago truncatula non-mycorrhizal and mycorrhizal roots. We identified 243 novel Medicago microRNAs and 118 mRNA cleavage targets of miRNA mature and star sequences. Several AM symbiosis-relevant genes were identified as miRNA targets. The transcript of MtNsp2, encoding a GRAS transcription factor involved in the nodule and mycorrhizal signaling pathway, is cleaved by a novel member of the miR171 gene family, namely miR171h. Here, we carried out a detailed analysis of the genomic structure of the MIR171h gene comprising our deep sequencing data. The results suggest a feedback circuit between mature miR171h and its own primary transcript showing the ability of this miRNA regulating itself. [less ▲]
Stars and symbiosis: microRNA- and microRNA*-mediated transcript cleavage involved in arbuscular mycorrhizal symbiosis.
Devers, Emanuel A.; Branscheid, Anja; May, Patrick et al
in Plant Physiology (2011), 156(4), 1990-2010
The majority of plants are able to form the arbuscular mycorrhizal (AM) symbiosis in association with AM fungi. During symbiosis development, plant cells undergo a complex reprogramming resulting in ... [more ▼]
The majority of plants are able to form the arbuscular mycorrhizal (AM) symbiosis in association with AM fungi. During symbiosis development, plant cells undergo a complex reprogramming resulting in profound morphological and physiological changes. MicroRNAs (miRNAs) are important components of the regulatory network of plant cells. To unravel the impact of miRNAs and miRNA-mediated mRNA cleavage on root cell reprogramming during AM symbiosis, we carried out high-throughput (Illumina) sequencing of small RNAs and degradome tags of Medicago truncatula roots. This led to the annotation of 243 novel miRNAs. An increased accumulation of several novel and conserved miRNAs in mycorrhizal roots suggest a role of these miRNAs during AM symbiosis. The degradome analysis led to the identification of 185 root transcripts as mature miRNA and also miRNA*-mediated mRNA cleavage targets. Several of the identified miRNA targets are known to be involved in root symbioses. In summary, the increased accumulation of specific miRNAs and the miRNA-mediated cleavage of symbiosis-relevant genes indicate that miRNAs are an important part of the regulatory network leading to symbiosis development. [less ▲]
Modeling RNA loops using sequence homology and geometric constraints.
Schudoma, Christian; May, Patrick ; Walther, Dirk
in Bioinformatics (2010), 26(13), 1671-2
SUMMARY: RNA loop regions are essential structural elements of RNA molecules influencing both their structural and functional properties. We developed RLooM, a web application for homology-based modeling ... [more ▼]
SUMMARY: RNA loop regions are essential structural elements of RNA molecules influencing both their structural and functional properties. We developed RLooM, a web application for homology-based modeling of RNA loops utilizing template structures extracted from the PDB. RLooM allows the insertion and replacement of loop structures of a desired sequence into an existing RNA structure. Furthermore, a comprehensive database of loops in RNA structures can be accessed through the web interface. AVAILABILITY AND IMPLEMENTATION: The application was implemented in Python, MySQL and Apache. A web interface to the database and loop modeling application is freely available at http://rloom.mpimp-golm.mpg.de CONTACT: schudoma@mpimp-golm.mpg.de; may@mpimp-golm.mpg.de; walther@mpimp-golm.mpg.de [less ▲]
Algebraic connectivity may explain the evolution of gene regulatory networks.
Nikoloski, Zoran; May, Patrick ; Selbig, Joachim
in Journal of Theoretical Biology (2010), 267(1), 7-14
Gene expression is a result of the interplay between the structure, type, kinetics, and specificity of gene regulatory interactions, whose diversity gives rise to the variety of life forms. As the dynamic ... [more ▼]
Gene expression is a result of the interplay between the structure, type, kinetics, and specificity of gene regulatory interactions, whose diversity gives rise to the variety of life forms. As the dynamic behavior of gene regulatory networks depends on their structure, here we attempt to determine structural reasons which, despite the similarities in global network properties, may explain the large differences in organismal complexity. We demonstrate that the algebraic connectivity, the smallest non-trivial eigenvalue of the Laplacian, of the directed gene regulatory networks decreases with the increase of organismal complexity, and may therefore explain the difference between the variety of analyzed regulatory networks. In addition, our results point out that, for the species considered in this study, evolution favours decreasing concentration of strategically positioned feed forward loops, so that the network as a whole can increase the specificity towards changing environments. Moreover, contrary to the existing results, we show that the average degree, the length of the longest cascade, and the average cascade length of gene regulatory networks cannot recover the evolutionary relationships between organisms. Whereas the dynamical properties of special subnetworks are relatively well understood, there is still limited knowledge about the evolutionary reasons for the already identified design principles pertaining to these special subnetworks, underlying the global quantitative features of gene regulatory networks of different organisms. The behavior of the algebraic connectivity, which we show valid on gene regulatory networks extracted from curated databases, can serve as an additional evolutionary principle of organism-specific regulatory networks. [less ▲]
PTGL: a database for secondary structure-based protein topologies.
May, Patrick ; Kreuchwig, Annika; Steinke, Thomas et al
in Nucleic Acids Research (2010), 38(Database issue), 326-30
With growing amount of experimental data, the number of known protein structures also increases continuously. Classification of protein structures helps to understand relationships between protein ... [more ▼]
With growing amount of experimental data, the number of known protein structures also increases continuously. Classification of protein structures helps to understand relationships between protein structure and function. The main classification methods based on secondary structures are SCOP, CATH and TOPS, which all classify under different aspects, and therefore can lead to different results. We developed a mathematically unique representation of protein structure topologies at a higher abstraction level providing new aspects of classification and enabling for a fast search through the data. Protein Topology Graph Library (PTGL; http://ptgl.zib.de) aims at providing a database on protein secondary structure topologies, including search facilities, the visualization as intuitive topology diagrams as well as in the 3D structure, and additional information. Secondary structure-based protein topologies are represented uniquely as undirected labeled graphs in four different ways allowing for exploration under different aspects. The linear notations, and the 2D and 3D diagrams of each notation facilitate a deeper understanding of protein topologies. Several search functions for topologies and sub-topologies, BLAST search possibility, and links to SCOP, CATH and PDBsum support individual and large-scale investigation of protein structures. Currently, PTGL comprises topologies of 54,859 protein structures. Main structural patterns for common structural motifs like TIM-barrel or Jelly Roll are pre-implemented, and can easily be searched. [less ▲]
Sequence-structure relationships in RNA loops: establishing the basis for loop homology modeling.
Schudoma, Christian; May, Patrick ; Nikiforova, Viktoria et al
in Nucleic Acids Research (2010), 38(3), 970-80
The specific function of RNA molecules frequently resides in their seemingly unstructured loop regions. We performed a systematic analysis of RNA loops extracted from experimentally determined three ... [more ▼]
The specific function of RNA molecules frequently resides in their seemingly unstructured loop regions. We performed a systematic analysis of RNA loops extracted from experimentally determined three-dimensional structures of RNA molecules. A comprehensive loop-structure data set was created and organized into distinct clusters based on structural and sequence similarity. We detected clear evidence of the hallmark of homology present in the sequence-structure relationships in loops. Loops differing by <25% in sequence identity fold into very similar structures. Thus, our results support the application of homology modeling for RNA loop model building. We established a threshold that may guide the sequence divergence-based selection of template structures for RNA loop homology modeling. Of all possible sequences that are, under the assumption of isosteric relationships, theoretically compatible with actual sequences observed in RNA structures, only a small fraction is contained in the Rfam database of RNA sequences and classes implying that the actual RNA loop space may consist of a limited number of unique loop structures and conserved sequences. The loop-structure data sets are made available via an online database, RLooM. RLooM also offers functionalities for the modeling of RNA loop structures in support of RNA engineering and design efforts. [less ▲]
Targeted proteomics for Chlamydomonas reinhardtii combined with rapid subcellular protein fractionation, metabolomics and metabolic flux analyses.
Wienkoop, Stefanie; Weiss, Julia; May, Patrick et al
in Molecular Biosystems (2010), 6(6), 1018-31
In the era of fast genome sequencing a critical goal is to develop genome-wide quantitative molecular approaches. Here, we present a metaproteogenomic strategy to integrate proteomics and metabolomics ... [more ▼]
In the era of fast genome sequencing a critical goal is to develop genome-wide quantitative molecular approaches. Here, we present a metaproteogenomic strategy to integrate proteomics and metabolomics data for systems level analysis in the recently sequenced unicellular green algae Chlamydomonas reinhardtii. To achieve a representative proteome coverage we analysed different growth conditions with protein prefractionation and shotgun proteomics. For protein identification, different genome annotations as well as new gene model predictions with stringent peptide filter criteria were used. An overlapping proteome coverage of 25%, consistent for all databases, was determined. The data are stored in a public mass spectral reference database ProMEX (http://www.promexdb.org/home.shtml). A set of proteotypic peptides comprising Calvin cycle, photosynthetic apparatus, starch synthesis, glycolysis, TCA cycle, carbon concentrating mechanisms (CCM) and other pathways was selected from this database for targeted proteomics (Mass Western). Rapid subcellular fractionation in combination with targeted proteomics allowed for measuring subcellular protein concentrations in attomole per 1000 cells. From the same samples metabolite concentrations and metabolic fluxes by stable isotope incorporation were analyzed. Differences were found in the growth-dependent crosstalk of chloroplastidic and mitochondrial metabolism. A Mass Western survey of all detectable carbonic anhydrases partially involved in carbon-concentrating mechanism (CCM) revealed highest internal cell concentrations for a specific low-CO2-inducible mitochondrial CAH isoform. This indicates its role as one of the strongest CO2-responsive proteins in the crosstalk of air-adapted mixotrophic chloroplast and mitochondrial metabolism in Chlamydomonas reinhardtii. [less ▲]