Biomedical subjects

J M Carazo

Publications and source records attributed to J M Carazo.

At least 37 records · Page 2Linked to original sources

PubMed1998

Self-organizing tree-growing network for the classification of protein sequences.

The self-organizing tree algorithm (SOTA) was recently introduced to construct phylogenetic trees from biological sequences, based on the principles of Kohonen's self-organizing maps and on Fritzke's growing cell structures. SOTA is designed in such a way that the generation of new nodes can be stopped when the sequences assigned to a node are already above a certain similarity threshold. In this way a phylogenetic tree resolved at a high taxonomic level can be obtained. This capability is especially useful to classify sets of diversified sequences. SOTA was originally designed to analyze pre-aligned sequences. It is now adapted to be able to analyze patterns associated to the frequency of residues along a sequence, such as protein dipeptide composition and other n-gram compositions. In this work we show that the algorithm applied to these data is able to not only successfully construct phylogenetic trees of protein families, such as cytochrome c, triosephophate isomerase, and hemoglobin alpha chains, but also classify very diversified sequence data sets, such as a mixture of interleukins and their receptors.

Algorithms↗

PubMed1998

3D reconstruction in electron microscopy using ART with smooth spherically symmetric volume elements (blobs).

Algebraic reconstruction techniques (ART) are iterative procedures for solving systems of linear equations. They have been used in tomography to recover objects from their projections. In this work we apply an ART approach in which the basis functions used to describe the objects are not based on voxels, but are much smoother functions named "blobs". The data collection studied in this work follows the so-called "conical tilt geometry" that is commonly used in many applications of three-dimensional electron microscopy of biological macromolecules. The performance of ART with blobs is carefully compared with a currently well-known three dimensional (3D) reconstruction algorithm (weighted back projection) using a methodology which assigns a level of statistical significance to a claim of relative superiority of one algorithm over another for a particular task. The conclusion we reach is that ART with blobs produces high-quality reconstructions and is, in particular, superior to weighted backprojection in recovering features along the "vertical" direction. For the exact implementation recommended in this paper, the computational costs of ART are almost an order of magnitude smaller than those of WBP.

Image Processing, Computer-Assisted↗

PubMed1998

Self-organizing tree growing network for classifying amino acids.

UNLABELLED: A self-organizing tree growing neural network was applied to classify amino acids and amino acid exchange matrices. AVAILABILITY: SOTA, is freely available by anonymous FTP or at http://www.cnb.uam.es/bioinfo/Software/so ta. CONTACT: carazo@cnb.uam.es

Algorithms↗

PubMed1998

Computational space reduction and parallelization of a new clustering approach for large groups of sequences.

MOTIVATION: The explosive growth of the biological sequences databases stimulated by genome projects has modified the framework of several applications in the biological sequence analysis area. In most cases, this new scenario is characterized by studies on large sets of sequences, suggesting the need for effective and automatic methods for their clustering. A more effective clustering of the database could be followed by the application of common family analysis schemes to the groups so formed. RESULTS: In this work, we present a new strategy to reduce the computational cost associated with the clustering of large sets of sequences which are expected to contain several families. The strategy is based on the grouping of the sequences into families by using a dynamic threshold on a pairwise sequence similarity criterion. Routine clustering of large data sets can now be done very efficiently. The method developed here achieves a computational space reduction of about an order of magnitude over more traditional ones of all-versus-all comparisons. The outcome of this approach produces family groupings that reproduce closely already accepted biological results. Our work includes a parallel implementation for distributed memory multiprocessors with a dynamic scheduling strategy for performance optimization. AVAILABILITY: By anonymous ftp at ftp.ac.uma.es (/pub/ots/pCluster directory), or from our Web site http://www.cnb. uam.es/www/software/software_index.html CONTACT: ots@ac.uma.es

Algorithms↗

PubMed1998

New phylogenetic venues opened by a novel implementation of the DNAml algorithm.

SUMMARY: This note describes the parallelization of a key biological application, such as the construction of phylogenetic trees, using the DNAml program. A careful run-time analysis allowed us to propose an efficient parallelization that scaled very well to large problem sizes. AVAILABILITY: By anonymous ftp at ftp.ac.uma. es (/pub/ots/pDNAml) or from our web site http://www.cnb.uam. es/www/software/software_index.html CONTACT: ots@ac.uma.es

Algorithms↗

PubMed1997

The RepA protein of plasmid RSF1010 is a replicative DNA helicase.

The RepA protein of the mobilizable broad host range plasmid RSF1010 has a key function in its replication. RepA is one of the smallest known helicases. The protein forms a homohexamer of 29,896-Da subunits. A variety of methods were used to analyze the quaternary structure of RepA. Gel filtration and cross-linking experiments demonstrated the hexameric structure, which was confirmed by electron microscopy and image reconstruction. These results agree with recent data obtained from RepA crystals diffracting at 3.5-A resolution (Röleke, D., Hoier, H., Bartsch, C., Umbach, P., Scherzinger, E., Lurz, R., and Saenger, W. (1997) Acta Crystallogr. Sec. D 53, 213-216). The RepA helicase has 5' --> 3' polarity. As do most true replicative helicases, RepA prefers a tailed substrate with an unpaired 3'-tail mimicking a replication fork. Optimal unwinding activity was achieved at the remarkably low pH of 5.5. In the presence of Mg2+ (Mn2+) ions, the RepA activity is fueled by ATP, dATP, GTP, and dGTP and less efficiently by CTP and dCTP. UTP and dTTP are poor effectors. Nonhydrolyzable ATP analogues, ADP, and pyrophosphate inhibit the helicase activity, whereas inorganic phosphate does not. The presence of Escherichia coli single-stranded DNA-binding protein stimulates unwinding at physiological pH 2-3-fold, whereas the RSF1010 replicon-specific primase, RepB' protein, has no effect, either in the presence or in the absence of single-stranded DNA-binding protein.

Adenosine Triphosphate↗

PubMed1997

Six molecules of SV40 large T antigen assemble in a propeller-shaped particle around a channel.

The large T antigen of simian virus 40 (SV40) is a multifunctional regulatory protein, responsible for both the control of viral infection and the required alterations of cellular processes. T antigen is the only viral protein required for viral DNA replication. It binds specifically to the viral origin and as a helicase unwinds the SV40 DNA bidirectionally. The functional complex is a double hexameric oligomer. In the absence of DNA, but in the presence of ATP or a non-hydrolyzable analog, T antigen assembles into hexamers, which are active as a helicase when a partially single-stranded (3') entry site exists on the substrate. We have used negative staining electron microscopy, single particle image processing and three-dimensional reconstruction with a new algebraic reconstruction techniques (ART) algorithm to study the structure of these hexameric particles in the presence of different nucleotide cofactors (ATP, ADP, and the non-hydrolyzable analogs ATPgammaS and AMP-PNP). In every case a strong 6-fold structure was found, with the six density maxima arranged in a ring-like particle around a channel, and a well-defined vorticity. Because these structural features have recently been found in other prokaryotic helicases, they seem to be strongly related to the activity of the protein, which suggests a general functional model conserved through evolution.

Adenosine Diphosphate↗

PubMed1997

Three-dimensional reconstruction from reduced sets of very noisy images acquired following a single-axis tilt schema: application of a new three-dimensional reconstruction algorithm and objective comparison with weighted backprojection.

In this work we propose a reconstruction algorithm (ART with blobs) that has not been previously used in electron Tomography and we compare it with the standard method in the field (weighted back projection, WBP). We assume that only a limited set of very noisy images, collected around a single axis tilt, is available; which is a typical situation in Electron Tomography. In general, the reconstruction problem is underdetermined (due to the limited number of projections) and the data are inconsistent (due to the high level of noise). The evaluation of the results is performed in a rigorous way by a task-oriented approach which makes use of numerical observers. ART with blobs outperforms WBP for a number of key tasks. Results are presented both for simplified line integral data and for realistic simulations of macromolecular structures embedded in amorphous ice.

Algorithms↗

PubMed1997

Phylogenetic reconstruction using an unsupervised growing neural network that adopts the topology of a phylogenetic tree.

We propose a new type of unsupervised, growing, self-organizing neural network that expands itself by following the taxonomic relationships that exist among the sequences being classified. The binary tree topology of this neutral network, contrary to other more classical neural network topologies, permits an efficient classification of sequences. The growing nature of this procedure allows to stop it at the desired taxonomic level without the necessity of waiting until a complete phylogenetic tree is produced. This novel approach presents a number of other interesting properties, such as a time for convergence which is, approximately, a lineal function of the number of sequences. Computer simulation and a real example show that the algorithm accurately finds the phylogenetic tree that relates the data. All this makes the neural network presented here an excellent tool for phylogenetic analysis of a large number of sequences.

Algorithms↗

PubMed1996

Analysis of structural variability within two-dimensional biological crystals by a combination of patch averaging techniques and self organizing maps.

We study in this work the use of self organizing maps to analyze the structural variability that can be found along two-dimensional crystals of biological macromolecules. Small areas of the crystals, termed "patches" by previous researchers, are used to obtain local average images that are then used as the input of a Self Organizing Map. This procedure allows for a fast and accurate image classification. Multivariate Statistical Analysis is then used on the resulting code vectors producing a very condensed data representation. This methodology is applied to previously studied crystals of bacteriophage phi 29 p10 connector, finding a crystalline heterogeneity probably associated to multilayers in some areas of the crystal.

Bacillus Phages↗

PubMed1995

An image-processing approach to dotplots: an X-Window-based program for interactive analysis of dotplots derived from sequence and structural data.

We present an approach to the study of the relationships between biological sequences and structures applying image analysis methods to dotplots. We introduce a set of analytical tools based on different types of digital image-processing filters that are new within the context of dotplots. We have reformulated some of the usual approaches in dotplot analysis as mathematical operations on images within the framework of mathematical morphology. An X-Window-based implementation of this new approach has been developed and is available by anonymous FTP.

Data Interpretation, Statistical↗

PubMed1994

Pattern recognition and classification of images of biological macromolecules using artificial neural networks.

The goal of this work was to analyze an image data set and to detect the structural variability within this set. Two algorithms for pattern recognition based on neural networks are presented, one that performs an unsupervised classification (the self-organizing map) and the other a supervised classification (the learning vector quantization). The approach has a direct impact in current strategies for structural determination from electron microscopic images of biological macromolecules. In this work we performed a classification of both aligned but heterogeneous image data sets as well as basically homogeneous but otherwise rotationally misaligned image populations, in the latter case completely avoiding the typical reference dependency of correlation-based alignment methods. A number of examples on chaperonins are presented. The approach is computationally fast and robust with respect to noise. Programs are available through ftp.

Algorithms↗

PubMed1994

On an efficient parallelization of exhaustive sequence comparison algorithms on message passing architectures.

We present a new parallel computing approach to the case of exhaustive sequential sequence comparison algorithms on message-passing architectures. In this context a modification of guided self-scheduling as well as efficient buffering strategies are presented. We discuss two specific implementations, one on the Paramid parallel computer, and the other on a cluster of workstations running PVM. In both cases the parallel performance is higher than with any other method presented so far. The code is public domain and can be obtained by anonymous ftp at ftp.cnb.uam.es.

Algorithms↗

PubMed1992

Three-dimensional structure of T3 connector purified from overexpressing bacteria.

The bacteriophage T3 connector has been purified from overexpressed protein in Escherichia coli, harboring a plasmid containing the gene encoding p8 protein. The connector, which is composed of 12 copies of p8, has been crystallized in two-dimensional sheets and studied by electron microscopy from negatively stained specimens. A two-dimensional Fourier filtering and averaging procedure was performed with crystalline specimens. In addition, single particle averaging techniques were used with other preparations. The average images obtained from these two approaches gave similar results. A three-dimensional reconstruction from two-dimensional crystals of T3 connectors was obtained by collecting several sets of tilted views and using standard Fourier procedures. The resolution of the three-dimensional map was 1.65 nm. The reconstructed connector shows two main domains: a wider one with 12 small units in the periphery and with an external diameter of 14.9 nm, and a smaller one with 8.5 nm diameter. The height of the reconstructed connector has been determined to be around 8.5 nm. The reconstruction clearly shows an internal open channel running along the longitudinal axis of the particle and having an average diameter of 3.7 nm.

Capsid↗

PubMed1992

Detection, classification and 3D reconstruction of biological macromolecules on hypercube computers.

In this work we present results of the mapping on hypercube computers of some of the key steps involved in the procedure for 3D structural determination from transmission electron microscopy images. The goal is the introduction of parallel processing tools in the field of electron microscopy image processing. We show how the rich topology of the hypercube, combined with an efficient programming strategy, allows for order-of-magnitude increase in computational capacity for such time-consuming tasks as calculation of multidimensional FFT's, cross-correlation coefficients, fuzzy partitioning functionals and the filtered back-projection 3D reconstruction method.

Algorithms↗

PubMed1991

Electron microscopy study of GroEL chaperonin: different views of the aggregate appear as a function of cell growth temperature.

We have studied two members of the family of morphogenetic factors or chaperonins, the GroEL-like factors from Escherichia coli and Bacillus subtilis, in order to determine the possible structural basis of their related function in promoting the correct and efficient assembly of biological oligomers. The main objective of this work has been to study by transmission electron microscopy the possible changes that these factors may undergo when subjected to a number of different conditions such as changes in temperature in vivo and in pH in vitro. We applied both rotational and multivariate statistical analyses of single particles to images of GroEl-like aggregates from the two bacteria. The most striking result is the finding of two distinct "front views" of these aggregates, from both E. coli and B. subtilis. One view, which has not been described earlier, shows a sixfold symmetry and is most abundant at growing temperatures below 37 degrees C. After heat shock, a view showing seven morphological units becomes dominant. On the basis of our analysis it is clear that GroEL-like morphogenetic factors from two unrelated bacteria such as E. coli and B. subtilis present two distinct views: one sixfold and the other sevenfold. Their relative percentage of appearance is related to the temperature at which the cells were grown and also to the storage conditions (pH).

Bacillus subtilis↗

PubMed1990

Fuzzy sets-based classification of electron microscopy images of biological macromolecules with an application to ribosomal particles.

Pattern recognition methods based on the theory of fuzzy sets are tested for their ability to classify electron microscopy images of biological specimens. The concept of fuzzy sets was chosen for its ability to represent classes of objects that are vaguely described from the measured data. A number of partitional clustering algorithms and an extensive set of cluster-validity functionals (some already reported and some newly developed) have been applied to a test-data set and to two real-data sets of images. One of the real-data sets corresponded to images of the Escherichia coli 50S ribosomal subunits depleted of proteins L7/L12 and the other set to images of the E. coli 70S monosome in the range of overlap views. These two latter sets had been previously studied by another clustering methodology. The new results obtained by the application of fuzzy clustering techniques will be compared to those previously obtained and some conclusions about the consistency of these classifications will be drawn from this comparison.

Algorithms↗

PubMed1989

Three-dimensional reconstruction of the ribosome from Escherichia coli.

Three-dimensional image reconstruction has been applied to electron micrographs of noncrystalline, negatively stained ribosomes obtained from Escherichia coli. Several independent reconstructions all show an overall appearance resembling models that had been derived earlier by direct visual interpretation of electron micrographs. The reconstructed ribosomes show numerous structural details not recognized previously, some of which may be functionally significant. A large elongate cavity (approximately 8-nm long x 5-nm wide x 6-nm [maximal] deep) is present on the surface of the ribosome near the base of its stalk and is identifiable as a portion of a feature termed the interface canyon, which was detected in prior reconstructions of the large ribosomal subunit (Radermacher, M., T. Wagenknecht, A. Verschoor, and J. Frank. 1987. EMBO (Eur. Mol. Biol. Organ.) J. 6:1107-1114). On the back of the ribosome, near the base of the central protuberance, is a hole leading to the interface canyon, which likely represents an exit site for the elongating polypeptide produced during protein biosynthesis. The exposed portion of the interface canyon appears well suited to bind two tRNA molecules in a configuration that is consistent with biochemical and structural data on the mechanism of peptide bond biosynthesis.

Escherichia coli↗