NGSmethDB: a database for next-generation sequencing single-cytosine-resolution DNA methylation data

Next-generation sequencing (NGS) together with bisulphite conversion allows the generation of whole genome methylation maps at single-cytosine resolution. This allows studying the absence of methylation in a particular genome region over a range of tissues, the differential tissue methylation or the changes occurring along pathological conditions. However, no database exists fully addressing such requirements. We propose here NGSmethDB (http://bioinfo2.ugr.es/NGSmethDB/gbrowse/) for the storage and retrieval of methylation data derived from NGS. Two cytosine methylation contexts (CpG and CAG/CTG) are considered. Through a browser interface coupled to a MySQL backend and several data mining tools, the user can search for methylation states in a set of tissues, retrieve methylation values for a set of tissues in a given chromosomal region, or display the methylation of promoters among different tissues. NGSmethDB is currently populated with human, mouse and Arabidopsis data, but other methylomes will be incorporated through an automatic pipeline as soon as new data become available. Dump downloads for three coverage levels (1, 5 or 10 reads) are available. NGSmethDB will be useful for experimental researchers, as well as for bioinformaticians, who might use the data as input for further research

On insertion-deletion systems over relational words

We introduce a new notion of a relational word as a finite totally ordered set of positions endowed with three binary relations that describe which positions are labeled by equal data, by unequal data and those having an undefined relation between their labels. We define the operations of insertion and deletion on relational words generalizing corresponding operations on strings. We prove that the transitive and reflexive closure of these operations has a decidable membership problem for the case of short insertion-deletion rules (of size two/three and three/two). At the same time, we show that in the general case such systems can produce a coding of any recursively enumerable language leading to undecidabilty of reachability questions.Comment: 24 pages, 8 figure

Different Transport Pathways of Individual Precursor Proteins in Mitochondria

Transport of mitochondrial precursor proteins into mitochondria of Neurospora crassa was studied in a cellfree reconstituted system. Precursors were synthesized in a reticulocyte lysate programmed with Neurospora mRNA and transported into isolated mitochondria in the absence of protein synthesis. Uptake of the following precursors was investigated: apocytochrome c, ADP/ATP carrier and subunit 9 of the oligomycin-sensitive ATPase. Addition of high concentrations of unlabelled chemically prepared apocytochrome c (1–10 μM) inhibited the appearance in the mitochondrial of labelled cytochrome c synthesized in vitro because the unlabelled protein dilutes the labelled one and because the translocation system has a limited capacity [apparent V is 1–3 pmol × min−1× (mg mitochondrial protein)−1]. Concentrations of added apocytochrome c exceeding the concentrations of precursor proteins synthesized in vitro by a factor of about 104 did not inhibit the transfer of ADP/ATP carrier or ATPase subunit 9 into mitochondria. Carbonylcyanide m-chlorophenylhydrazone, an uncoupler of oxidative phosphorylation, inhibited transfer in vitro of ADP/ATP carrier and of ATPase subunit 9, but not of cytochrome c. These findings suggest that cytochrome c and the other two proteins have different import pathways into mitochondria. It can be inferred from the data presented that different 'receptors' on the mitochondrial surface mediate the specific recognition of precursor proteins by mitochondria as a first step in the transport process

Transport of Proteins into Mitochondria

The mitochondrial ADP/ATP carrier is an integral transmembrane protein of the inner membrane. It is synthesized on cytoplasmic ribosomes. Kinetic data suggested that this protein is transferred into mitochondria in a posttranslational manner. The following results provide further evidence for such a mechanism and provide information on its details. 1. In homologous and heterologous translation systems the newly synthesized ADP/ATP carrier protein is present in the postribosomal supernatant. 2. Analysis by density gradient centrifugation and gel filtration shows, that the ADP/ATP carrier molecules in the postribosomal fraction are present as soluble complexes with apparent molecular weights of about 120000 and 500000 or larger. The carrier binds detergents such as Triton X-100 and deoxycholate forming mixed micelles with molecular weights of about 200000–400000. 3. Incubation of a postribosomal supernatant of a reticulocyte lysate containing newly synthesized ADP/ATP carrier with mitochondria isolated from Neurospora spheroplasts results in efficient transfer of the carrier into mitochondria. About 20–30% of the transferred carrier are resistant to proteinase in whole mitochondria. The authentic mature protein is also largely resistant to proteinase in whole mitochondria and sensitive after lysis of mitochondria with detergent. Integrity of mitochondria is a prerequisite for translocation into proteinase resistant position. 4. The transfer in vitro into a proteinase-resistant form is inhibited by the uncoupler carbonyl-cyanide m-chlorophenylhydrazone but not the proteinase-sensitive binding. These observations suggest that the posttranslational transfer of ADP/ATP carrier occurs via the cytosolic space through a soluble oligomeric precursor form. This precursor is taken up by intact mitochondria into an integral position in the membrane. These findings are considered to be of general importance for the intracellular transfer of insoluble membrane proteins. They support the view that such proteins can exist in a water-soluble form its precursors and upon integration into the membrane undergo a conformational change. Uptake into the membrane may involve the cleavage of an additional sequence in some proteins, but this appears not to be a prerequisite as demonstrated by the ADP/ATP carrier protein

Cell-Free Synthesis of the Mitochondrial ADP/ATP Carrier Protein of Neurospora crassa

ADP/ATP carrier protein was synthesized in heterologous cell-free systems programmed with Neurospora poly(A)-containing RNA and homologous cell-free systems from Neurospora. The apparent molecular weight of the product obtained in vitro was the same as that of the authentic mitochondrial protein. The primary translation product obtained in reticulocyte lysates starts with formylmethionine when formylated initiator methionyl-tRNA (fMet-tRNAfMet) was present. The product synthesized in vitro was released from the ribosomes into the postribosomal supernatant. The evidence presented indicates that the ADP/ATP carrier is synthesized as a polypeptide with the same molecular weight as the mature monomeric protein and does not carry an additional sequence

ContDist: a tool for the analysis of quantitative gene and promoter properties

<p>Abstract</p> <p>Background</p> <p>The understanding of how promoter regions regulate gene expression is complicated and far from being fully understood. It is known that histones' regulation of DNA compactness, DNA methylation, transcription factor binding sites and CpG islands play a role in the transcriptional regulation of a gene. Many high-throughput techniques exist nowadays which permit the detection of epigenetic marks and regulatory elements in the promoter regions of thousands of genes. However, so far the subsequent analysis of such experiments (e.g. the resulting gene lists) have been hampered by the fact that currently no tool exists for a detailed analysis of the promoter regions.</p> <p>Results</p> <p>We present ContDist, a tool to statistically analyze quantitative gene and promoter properties. The software includes approximately 200 quantitative features of gene and promoter regions for 7 commonly studied species. In contrast to "traditionally" ontological analysis which only works on qualitative data, all the features in the underlying annotation database are quantitative gene and promoter properties.</p> <p>Utilizing the strong focus on the promoter region of this tool, we show its usefulness in two case studies; the first on differentially methylated promoters and the second on the fundamental differences between housekeeping and tissue specific genes. The two case studies allow both the confirmation of recent findings as well as revealing previously unreported biological relations.</p> <p>Conclusion</p> <p>ContDist is a new tool with two important properties: 1) it has a strong focus on the promoter region which is usually disregarded by virtually all ontology tools and 2) it uses quantitative (continuously distributed) features of the genes and its promoter regions which are not available in any other tool. ContDist is available from <url>http://web.bioinformatics.cicbiogune.es/CD/ContDistribution.php</url></p

Sagittal jaw position in relation to body posture in adult humans – a rasterstereographic study

BACKGROUND: The correlations between the sagittal jaw position and the cranio – cervical inclination are described in literature. Only few studies focus on the sagittal jaw position and the body posture using valid and objective orthopaedic examination methods. The aim of this study was to test the hypothesis that patients with malocclusions reveal significant differences in body posture compared to those without (upper thoracic inclination, kyphotic angle, lordotic angle and lower lumbar inclination). METHODS: Eighty-four healthy adult patients (with a mean age = 25.6 years and ranging from 16.1 to 55.8 years) were examined with informed consent. The orthodontic examination horizontal overjet (distance between upper and lower incisors) was determined by using an orthodontic digital sliding calliper. The subjects were subdivided in respect of the overjet with the following results: 18 revealed a normal overjet (Class I), 38 had an increased overjet (Class II) and 28 had an reversed overjet (Class III). Rasterstereography was used to carry out a three – dimensional back shape analysis. This method is based on photogrammetry. A three-dimensional shape was produced by analysing the distortion of parallel horizontal white light lines projected on the patient's back, followed by mathematical modelling. On the basis of the sagittal profile the upper thoracic inclination, the thoracic angle, the lordotic angle and the pelvic inclination were determined with a reported accuracy of 2.8° and the correlations to the sagittal jaw position were calculated by means of ANOVA, Scheffé and Kruskal-Wallis procedures. RESULTS: Between the different overjet groups, no statistically significant differences or correlations regarding the analysed back shape parameters could be obtained. However, comparing males and females there were statistically significant differences in view of the parameters 'lordotic angle' and 'pelvic inclination'. CONCLUSION: No correlations between overjet and variables of the thoracic, lordotic or the pelvic inclination could be observed

Molecular gas in the galaxy cluster Abell 262. CO observations of UGC 1347 and other galaxies of the cluster

We present millimeter CO line emission observations of 12 galaxies within the Abell 262 cluster, together with L_FIR data, in the context of a possible molecular gas deficiency within the region of the cluster center. Several indications of the presence of such a deficiency are highlighted and connected to a model of cirrus-like cloud stripping. The model predicts a drop in the average 100 micron flux density of galaxies in the core of the cluster compared to the average 100 micron flux density in the outer regions, which is actually indicated in the IRAS data of the cluster members. This drop is explained by the decrease in the total hydrogen column density N(H) and, therefore, also includes a decrease in the molecular gas content. In addition to results for the global CO content of the galaxy sample, high-resolution interferometric CO(1-0) observations of one of the cluster members, UGC 1347, exemplify the spatial distribution of the molecular gas in a galaxy of the cluster. With these observations, it was possible to confirm the existence of a bright off-nuclear CO-emission source and to derive molecular masses and line ratios for this source and the nucleus.Comment: 14 pages, 14 figures, accepted by A&