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PubMed2006

Functional annotation of the Arabidopsis P450 superfamily based on large-scale co-expression analysis.

Cytochrome P450 mono-oxygenases play prominent roles in a diverse set of metabolic pathways, but the function of most of these enzymes remains obscure. A bottleneck in the functional genomics of this superfamily constitutes hypothesis generation to identify potential substrates (or substrate classes) individual P450s may act on. We used publicly available large-scale expression data to perform co-expression analysis comparing the expression matrix of each P450 with those from more than 4000 selected genes across thousands of microarrays. Based on functional annotations of co-expressed genes from a diverse set of databases, co-expressed pathways were thus identified for each P450. Using this approach, most P450s with known functions were placed into their respective pathways, thereby proofing the concept. As examples, pathway mapping results identifying novel P450s potentially acting on flower-specific monoterpenes and root-specific triterpenes are described. Co-expression results for all Arabidopsis P450s will be presented as a web resource on the 'CYPedia' web pages (http://ibmp.u-strasbg.fr/CYPedia/).

Arabidopsis↗

PubMed2002

Functional annotation of mammalian genomic DNA sequence by chemical mutagenesis: a fine-structure genetic mutation map of a 1- to 2-cM segment of mouse chromosome 7 corresponding to human chromosome 11p14-p15.

Eleven independent, recessive, N-ethyl-N-nitrosourea-induced mutations that map to a approximately 1- to 2-cM region of mouse chromosome (Chr) 7 homologous to human Chr 11p14-p15 were recovered from a screen of 1,218 gametes. These mutations were initially identified in a hemizygous state opposite a large p-locus deletion and subsequently were mapped to finer genomic intervals by crosses to a panel of smaller p deletions. The 11 mutations also were classified into seven complementation groups by pairwise crosses. Four complementation groups were defined by seven prenatally lethal mutations, including a group (l7R3) comprised of two alleles of obvious differing severity. Two allelic mutations (at the psrt locus) result in a severe seizure and runting syndrome, but one mutation (at the fit2 locus) results in a more benign runting phenotype. This experiment has added seven loci, defined by phenotypes of presumed point mutations, to the genetic map of a small (1-2 cM) region of mouse Chr 7 and will facilitate the task of functional annotation of DNA sequence and transcription maps both in the mouse and the corresponding human 11p14-p15 homology region.

Animals↗

PubMed2000

Comparative organization of cattle chromosome 5 revealed by comparative mapping by annotation and sequence similarity and radiation hybrid mapping.

A whole genome cattle-hamster radiation hybrid cell panel was used to construct a map of 54 markers located on bovine chromosome 5 (BTA5). Of the 54 markers, 34 are microsatellites selected from the cattle linkage map and 20 are genes. Among the 20 mapped genes, 10 are new assignments that were made by using the comparative mapping by annotation and sequence similarity strategy. A LOD-3 radiation hybrid framework map consisting of 21 markers was constructed. The relatively low retention frequency of markers on this chromosome (19%) prevented unambiguous ordering of the other 33 markers. The length of the map is 398.7 cR, corresponding to a ratio of approximately 2.8 cR(5,000)/cM. Type I genes were binned for comparison of gene order among cattle, humans, and mice. Multiple internal rearrangements within conserved syntenic groups were apparent upon comparison of gene order on BTA5 and HSA12 and HSA22. A similarly high number of rearrangements were observed between BTA5 and MMU6, MMU10, and MMU15. The detailed comparative map of BTA5 should facilitate identification of genes affecting economically important traits that have been mapped to this chromosome and should contribute to our understanding of mammalian chromosome evolution.

Animals↗

PubMed2003

Functional annotation of the putative orphan Caenorhabditis elegans G-protein-coupled receptor C10C6.2 as a FLP15 peptide receptor.

This report describes the cloning and functional annotation of a Caenorhabditis elegans orphan G-protein-coupled receptor (GPCR) (C10C6.2) as a receptor for the FMRFamide-related peptides (FaRPs) encoded on the flp15 precursor gene, leading to the receptor designation FLP15-R. A cDNA encoding C10C6.2 was obtained using PCR techniques, confirmed identical to the Worm-pep-predicted sequence, and cloned into a vector appropriate for eucaryotic expression. A [35S]guanosine 5'-O-(thiotriphosphate) (GTPgammaS) assay with membranes prepared from Chinese hamster ovary (CHO) cells transiently transfected with FLP15-R was used as a read-out for receptor activation. FLP15-R was activated by putative FLP15 peptides, GGPQGPLRF-NH2 (FLP15-1), RGPSGPLRF-NH2 (FLP15-2A), its des-Arg1 counterpart, GPSGPLRF-NH2 (FLP15-2B), and to a lesser extent, by a tobacco hornworm Manduca sexta FaRP, GNSFLRFNH2 (F7G) (potency ranking FLP15-2A > FLP15-1 > FLP15-2B >> F7G). FLP15-R activation was abolished in the transfected cells pretreated with pertussis toxin, suggesting a preferential receptor coupling to Gi/Go proteins. The functional expression of FLP15-R in mammalian cells was temperature-dependent. Either no stimulation or significantly lower ligand-evoked [35S]GTPgammaS binding was observed in membranes prepared from transfected FLP15-R/CHO cells cultured at 37 degrees C. However, a 37 to 28 degrees C temperature shift implemented 24 h post-transfection consistently resulted in an improved activation signal and was essential for detectable functional expression of FLP15-R in CHO cells. To our knowledge, the FLP15 receptor is only the second deorphanized C. elegans neuropeptide GPCR reported to date.

Amino Acid Sequence↗

PubMed2005

Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth.

Here we describe a proteomic analysis of Escherichia coli in which 3,199 protein forms were detected, and of those 2,160 were annotated and assigned to the cytosol, periplasm, inner membrane, and outer membrane by biochemical fractionation followed by two-dimensional gel electrophoresis and tandem mass spectrometry. Represented within this inventory were unique and modified forms corresponding to 575 different ORFs that included 151 proteins whose existence had been predicted from hypothetical ORFs, 76 proteins of completely unknown function, and 222 proteins currently without location assignments in the Swiss-Prot Database. Of the 575 unique proteins identified, 42% were found to exist in multiple forms. Using DIGE, we also examined the relative changes in protein expression when cells were grown in the presence and absence of amino acids. A total of 23 different proteins were identified whose abundance changed significantly between the two conditions. Most of these changes were found to be associated with proteins involved in carbon and amino acid metabolism, transport, and chemotaxis. Detailed information related to all 2,160 protein forms (protein and gene names, accession numbers, subcellular locations, relative abundances, sequence coverage, molecular masses, and isoelectric points) can be obtained upon request in either tabular form or as interactive gel images.

Amino Acids↗

PubMed2004

Two novel LEM-domain proteins are splice products of the annotated Drosophila melanogaster gene CG9424 (Bocksbeutel).

The LEM motif is a sequence of 40-50 amino acids that has been identified in a number of non-related proteins of the inner nuclear membrane including the lamina-associated polypeptides 2 (LAP2), emerin, MAN1 and the Drosophila protein otefin. This evolutionary conserved sequence motif can mediate via the interaction with the small protein BAF the binding of LEM-domain proteins to DNA. Taking advantage of its sequenced genome we analyzed whether Drosophila possesses beside otefin additional genes coding for proteins with a LEM motif. A putative candidate gene was the annotated gene CG9424 which we named Bocksbeutel. Of all putative Drosophila LEM-domain proteins, otefin and Bocksbeutel exhibited the highest similarity in the LEM motif (53% identical amino acids). The Bocksbeutel gene can code for two isoforms of 399 and 351 amino acids that are produced by alternative splicing. In the alpha-isoform a transmembrane domain is localized close to the carboxyterminus. This segment is absent in the shorter beta-isoform. By RT-PCR we could show that in the embryo the mRNA coding for the alpha-isoform and in significantly lower amounts the mRNA coding for the beta-isoform are expressed. When expressed in transfected cells as GFP fusion proteins, the beta-isoform is localized predominantly in the nucleoplasm and the alpha-isoform is targeted to the nuclear envelope, indicating that Bocksbeutel-alpha is localized in the inner nuclear membrane. Bocksbeutel-alpha is the predominant isoform expressed in cells, larvae, and flies. Indirect immunofluorescence with Bocksbeutel-specific antibodies on tissues and cultured cells revealed that Bocksbeutel proteins are localized in the nuclear envelope and in the cytoplasm. By RNA interference we have down-regulated the expression of Bocksbeutel, BAF, otefin, and lamin DmO in Drosophila Kc167 cells. The down-regulation of Bocksbeutel and otefin had no influence on the viability of Kc167 cells and the intracellular localization of all other nuclear and nuclear envelope proteins analyzed. In contrast, when lamin DmO was reduced by RNAi the distribution of Bocksbeutel and otefin in the nuclear envelope of Kc167 cells was significantly altered. We conclude that the two LEM-domain proteins Bocksbeutel and otefin are no limiting components for the maintenance of the nuclear architecture in cultured Drosophila cells at interphase.

Alternative Splicing↗

PubMedDate not supplied

Children's health promotion through caregiver preparation in pediatric brain injury settings: compensating for shortened hospital stays with a three-phase model of health education and annotated bibliography.

Shorter hospital and rehabilitation stays in cases of pediatric brain injury necessitate greater preparation and training of family caregivers, who often will be responsible for complex and continued care at home. At the same time, a growing nursing shortage results in less available time for individualized, one-on-one caregiver education in medical settings prior to discharge. What is needed are innovative models of caregiver preparation and education that are comprehensive, systematic, and maximize the use of health professionals' limited time. The model presented here aims to provide a progressive three-phase model of caregiver education that makes efficient use of health professionals' time and delivers crucial information in a time-released manner throughout the entire continuum of care under the guidance of health professionals. An annotated bibliography of published caregiver education resources and the appropriate time for their delivery to family members is provided in the Appendix.

Bibliographies as Topic↗

PubMed2006

A select annotated bibliography: illegal drug research in rural and suburban areas.

As the diffusion of illegal drugs continues to spread to rural and suburban areas, there is a greater need among illegal drug researchers, law enforcement, and policy makers to gain knowledge from previous work done on what is loosely termed as rural substance abuse research. To help serve that need an annotated bibliography is proffered based on three categories and one method of illegal substance abuse research. These are: drug consumption, drug distribution, drug prices, and ethnography. An exhaustive review of the literature was not undertaken, but rather a focus on research that addresses drug consumption and distribution is put forward.

Humans↗

PubMed1993

Medical education for hospice care: a selected bibliography with brief annotations.

This is a briefly annotated bibliography of useful materials for the education of health professionals, principally physicians. It encompasses teaching goals, methods, and settings, as well as model courses, course evaluation, communication skills, general resources on death education, and miscellaneous background pieces.

Education, Medical↗

PubMedDate not supplied

Nontrauma helicopter emergency medical services transport: annotated review of selected outcomes-related literature.

While helicopter emergency medical services (HEMS) has its roots in military transport of wounded soldiers, rotor-wing transport is also used for a wide variety of nontrauma indications. Despite this common use of HEMS for noninjured patients, a Medline search found little systematic review of the literature pertinent to HEMS use for nontrauma. With HEMS utilization subject to appropriately increased scrutiny, those seeking to research HEMS utility in noninjured patients could benefit from existence of a collection of the topical literature. This paper aims to provide such a review, in the form of an annotated bibliography of Index Medicus journal studies assessing potential medical risks and benefits of HEMS transport for noninjured patients. The paper's goal is to provide a useful resource for those interested in pursuing more focused review of various sectors of the nontrauma HEMS literature. As such, the main objective of the article summaries is to provide a brief outline of study design and results; there is also limited editorial comment included after each summary.

Air Ambulances↗

PubMed1994

The accuracy of power-spectrum analysis of heart-rate variability from annotated RR lists generated by Holter systems.

The accuracy of spectral analysis of heart-rate variability performed on annotated RR interval lists obtained from several commercial Holter systems was appraised. Five tape-recorder-based systems (Del Mar 750, Marquette 8000, Oxford Medilog Excel, Remco Cardioline AD 35 and Reynolds Pathfinder PA3) and four solid-state systems (Hewlett Packard 43420B, Marquette Seer, Oxford 6000FD2, Reynolds E-Ram) were considered. Two ECG signals with fixed real morphology but characterized by a different degree of modulation of the RR interval (reduced and normal variability) were fed into the recorders evaluated. The total power and the power in the very low-, low- and high-frequency bands were then estimated on all Holter-generated RR sequences. Spectral analysis was performed by both the autoregressive and fast-Fourier-transform methods. The estimation error of each parameter was statistically characterized and, for tape-recorder-based systems, inferential analysis was used to test for differences between recorders, tapes and times of recording. The centre and dispersion of the estimation error changed markedly from system to system. Some tape-recording systems showed large inter-recorder differences. The degree of spectral distortion was never uniform among selected bands. Solid-state systems performed better than tape-recording ones but both were limited in the accuracy by the quantization of RR interval measurement. The fast Fourier method yielded spectral estimates more stable than the autoregressive method. Our data clearly show that spectral analysis of very low-variability signals may be seriously affected by Holter recording and preprocessing of ECG signals.

Electrocardiography↗

PubMedDate not supplied

A dictionary-based approach for gene annotation.

This paper describes a fast and fully automated dictionary-based approach to gene annotation and exon prediction. Two dictionaries are constructed, one from the nonredundant protein OWL database and the other from the dbEST database. These dictionaries are used to obtain O (1) time lookups of tuples in the dictionaries (4 tuples for the OWL database and 11 tuples for the dbEST database). These tuples can be used to rapidly find the longest matches at every position in an input sequence to the database sequences. Such matches provide very useful information pertaining to locating common segments between exons, alternative splice sites, and frequency data of long tuples for statistical purposes. These dictionaries also provide the basis for both homology determination, and statistical approaches to exon prediction.

Alternative Splicing↗

PubMed2001

Annotated draft genomic sequence from a Streptococcus pneumoniae type 19F clinical isolate.

The public availability of numerous microbial genomes is enabling the analysis of bacterial biology in great detail and with an unprecedented, organism-wide and taxon-wide, broad scope. Streptococcus pneumoniae is one of the most important bacterial pathogens throughout the world. We present here sequences and functional annotations for 2.1-Mbp of pneumococcal DNA, covering more than 90% of the total estimated size of the genome. The sequenced strain is a clinical isolate resistant to macrolides and tetracycline. It carries a type 19F capsular locus, but multilocus sequence typing for several conserved genetic loci suggests that the strain sequenced belongs to a pneumococcal lineage that most often expresses a serotype 15 capsular polysaccharide. A total of 2,046 putative open reading frames (ORFs) longer than 100 amino acids were identified (average of 1,009 bp per ORF), including all described two-component systems and aminoacyl tRNA synthetases. Comparisons to other complete, or nearly complete, bacterial genomes were made and are presented in a graphical form for all the predicted proteins.

DNA, Bacterial↗

PubMed2006

On the repeat-annotated phylogenetic tree reconstruction problem.

A new problem in phylogenetic inference is presented, based on recent biological findings indicating a strong association between reversals (i.e., inversions) and repeats. These biological findings are formalized here in a new mathematical model, called repeat-annotated phylogenetic trees (RAPT). We show that, under RAPT, the evolutionary process--including both the tree-topology as well as internal node genome orders--is uniquely determined, a property that is of major significance both in theory and in practice. Furthermore, the repeats are employed to provide linear-time algorithms for reconstructing both the genomic orders and the phylogeny, which are NP-hard problems under the classical model of sorting by reversals (SBR).

Algorithms↗

PubMed2006

Combining gene annotations and gene expression data in model-based clustering: weighted method.

It has been increasingly recognized that incorporating prior knowledge into cluster analysis can result in more reliable and meaningful clusters. In contrast to the standard modelbased clustering with a global mixture model, which does not use any prior information, a stratified mixture model was recently proposed to incorporate gene functions or biological pathways as priors in model-based clustering of gene expression profiles: various gene functional groups form the strata in a stratified mixture model. Albeit useful, the stratified method may be less efficient than the global analysis if the strata are non-informative to clustering. We propose a weighted method that aims to strike a balance between a stratified analysis and a global analysis: it weights between the clustering results of the stratified analysis and that of the global analysis; the weight is determined by data. More generally, the weighted method can take advantage of the hierarchical structure of most existing gene functional annotation systems, such as MIPS and Gene Ontology (GO), and facilitate choosing appropriate gene functional groups as priors. We use simulated data and real data to demonstrate the feasibility and advantages of the proposed method.

Algorithms↗

PubMed1998

EUCLID: automatic classification of proteins in functional classes by their database annotations.

UNLABELLED: A tool is described for the automatic classification of sequences in functional classes using their database annotations. The Euclid system is based on a simple learning procedure from examples provided by human experts. AVAILABILITY: Euclid is freely available for academics at http://www.gredos.cnb.uam.es/EUCLID, with the corresponding dictionaries for the generation of three, eight and 14 functional classes. CONTACT: E-mail: valencia@cnb.uam.es SUPPLEMENTARY INFORMATION: The results of the EUCLID classification of different genomes are available at http://www.sander.ebi.ac. uk/genequiz/. A detailed description of the different applications mentioned in the text is available at http://www.gredos.cnb.uam. es/EUCLID/Full_Paper

Computational Biology↗

PubMed1999

TargetFinder: searching annotated sequence databases for target genes of transcription factors.

UNLABELLED: TargetFinder is a new software tool to search a database of annotated sequences for transcription factor binding sites located in context with other important transcription regulatory signals and regions, like the TATA element, the promoter, and so on, thereby greatly reducing the background usually associated with this kind of search. AVAILABILITY: The TargetFinder Web service is available at http://hercules.tigem.it/TargetFinder.html CONTACT: giovanni.lavorgna@hsr.it

Binding Sites↗

PubMed2000

SAWTED: structure assignment with text description--enhanced detection of remote homologues with automated SWISS-PROT annotation comparisons.

MOTIVATION: Sequence database search methods often identify putative sub-threshold hits of known function or structure for a given query sequence. It is widespread practice to filter these hits by hand using knowledge of function and other factors; to the expert, some hits may appear more sensible than others. SAWTED (Structure Assignment With Text Description) is an automated solution to this post-filtering problem which will be applicable to large scale genome assignments. RESULTS: A standard document comparison algorithm is applied to text descriptions extracted from SWISS-PROT annotations. The added value of SAWTED in combination with PSI-BLAST has been shown with a benchmark of difficult remote homologues taken from the SCOP structure database. AVAILABILITY: A WAWTED PSI-BLAST Web server is available to perform sensitive searches against the protein structure database (http://www.bmm.icnet.uk/servers/sawted). CONTACT: R.MacCallum@icrf.icnet.uk

Algorithms↗