42,888 research outputs found
5SRNAdb: an information resource for 5S ribosomal RNAs
Ribosomal 5S RNA (5S rRNA) is the ubiquitous RNA component found in the large
subunit of ribosomes in all known organisms. Due to its small size, abundance
and evolutionary conservation 5S rRNA for many years now is used as a model
molecule in studies on RNA structure, RNAâprotein interactions and molecular
phylogeny. 5SRNAdb (http://combio.pl/5srnadb/) is the first database that
provides a high quality reference set of ribosomal 5S RNAs (5S rRNA) across
three domains of life. Here, we give an overview of new developments in the
database and associated web tools since 2002, including updates to database
content, curation processes and user web interfaces
A creature with a hundred waggly tails: intrinsically disordered proteins in the ribosome
This article is made available for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.Intrinsic disorder (i.e., lack of a unique 3-D structure) is a common phenomenon, and many biologically active proteins are disordered as a whole, or contain long disordered regions. These intrinsically disordered proteins/regions constitute a significant part of all proteomes, and their functional repertoire is complementary to functions of ordered proteins. In fact, intrinsic disorder represents an important driving force for many specific functions. An illustrative example of such disorder-centric functional class is RNA-binding proteins. In this study, we present the results of comprehensive bioinformatics analyses of the abundance and roles of intrinsic disorder in 3,411 ribosomal proteins from 32 species. We show that many ribosomal proteins are intrinsically disordered or hybrid proteins that contain ordered and disordered domains. Predicted globular domains of many ribosomal proteins contain noticeable regions of intrinsic disorder. We also show that disorder in ribosomal proteins has different characteristics compared to other proteins that interact with RNA and DNA including overall abundance, evolutionary conservation, and involvement in proteinâprotein interactions. Furthermore, intrinsic disorder is not only abundant in the ribosomal proteins, but we demonstrate that it is absolutely necessary for their various functions
Representation, searching and discovery of patterns of bases in complex RNA structures
We describe a graph theoretic method designed to perform efficient searches for substructural patterns in nucleic acid structural coordinate databases using a simplified vectorial representation. Two vectors represent each nucleic acid base and the relative positions of bases with respect to one another are described in terms of distances between the defined start and end points of the vectors on each base. These points comprise the nodes and the distances the edges of a graph, and a pattern search can then be performed using a subgraph isomorphism algorithm. The minimal representation was designed to facilitate searches for complex patterns but was first tested on simple, well-characterised arrangements of bases such as base pairs and GNRA-tetraloop receptor interactions. The method performed very well for these interaction types. A survey of side-by-side base interactions, of which the adenosine platform is the best known example, also locates examples of similar base rearrangements that we consider to be important in structural regulation. A number of examples were found, with GU platforms being particularly prevalent. A GC platform in the RNA of the Thermus thermophilus small ribosomal subunit is in an analogous position to an adenosine platform in other species. An unusual GG platform is also observed close to one of the substrate binding sites in Haloarcula marismortui large ribosomal subunit RNA
Size, shape, and flexibility of RNA structures
Determination of sizes and flexibilities of RNA molecules is important in
understanding the nature of packing in folded structures and in elucidating
interactions between RNA and DNA or proteins. Using the coordinates of the
structures of RNA in the Protein Data Bank we find that the size of the folded
RNA structures, measured using the radius of gyration, , follows the Flory
scaling law, namely, \AA where N is the number of
nucleotides. The shape of RNA molecules is characterized by the asphericity
and the shape parameters that are computed using the eigenvalues
of the moment of inertia tensor. From the distribution of , we find
that a large fraction of folded RNA structures are aspherical and the
distribution of values shows that RNA molecules are prolate (). The
flexibility of folded structures is characterized by the persistence length
. By fitting the distance distribution function to the worm-like
chain model we extracted the persistence length . We find that \AA. The dependence of on implies the average length of
helices should increases as the size of RNA grows. We also analyze packing in
the structures of ribosomes (30S, 50S, and 70S) in terms of , ,
, and . The 70S and the 50S subunits are more spherical compared to
most RNA molecules. The globularity in 50S is due to the presence of an
unusually large number (compared to 30S subunit) of small helices that are
stitched together by bulges and loops. Comparison of the shapes of the intact
70S ribosome and the constituent particles suggests that folding of the
individual molecules might occur prior to assembly.Comment: 28 pages, 8 figures, J. Chem. Phys. in pres
Systematic identification of gene families for use as markers for phylogenetic and phylogeny- driven ecological studies of bacteria and archaea and their major subgroups
With the astonishing rate that the genomic and metagenomic sequence data sets
are accumulating, there are many reasons to constrain the data analyses. One
approach to such constrained analyses is to focus on select subsets of gene
families that are particularly well suited for the tasks at hand. Such gene
families have generally been referred to as marker genes. We are particularly
interested in identifying and using such marker genes for phylogenetic and
phylogeny-driven ecological studies of microbes and their communities. We
therefore refer to these as PhyEco (for phylogenetic and phylogenetic ecology)
markers. The dual use of these PhyEco markers means that we needed to develop
and apply a set of somewhat novel criteria for identification of the best
candidates for such markers. The criteria we focused on included universality
across the taxa of interest, ability to be used to produce robust phylogenetic
trees that reflect as much as possible the evolution of the species from which
the genes come, and low variation in copy number across taxa. We describe here
an automated protocol for identifying potential PhyEco markers from a set of
complete genome sequences. The protocol combines rapid searching, clustering
and phylogenetic tree building algorithms to generate protein families that
meet the criteria listed above. We report here the identification of PhyEco
markers for different taxonomic levels including 40 for all bacteria and
archaea, 114 for all bacteria, and much more for some of the individual phyla
of bacteria. This new list of PhyEco markers should allow much more detailed
automated phylogenetic and phylogenetic ecology analyses of these groups than
possible previously.Comment: 24 pages, 3 figure
Transcription-translation coupling: direct interactions of RNA polymerase with ribosomes and ribosomal subunits.
In prokaryotes, RNA polymerase and ribosomes can bind concurrently to the same RNA transcript, leading to the functional coupling of transcription and translation. The interactions between RNA polymerase and ribosomes are crucial for the coordination of transcription with translation. Here, we report that RNA polymerase directly binds ribosomes and isolated large and small ribosomal subunits. RNA polymerase and ribosomes form a one-to-one complex with a micromolar dissociation constant. The formation of the complex is modulated by the conformational and functional states of RNA polymerase and the ribosome. The binding interface on the large ribosomal subunit is buried by the small subunit during protein synthesis, whereas that on the small subunit remains solvent-accessible. The RNA polymerase binding site on the ribosome includes that of the isolated small ribosomal subunit. This direct interaction between RNA polymerase and ribosomes may contribute to the coupling of transcription to translation
Two RNA-binding motifs in eIF3 direct HCV IRES-dependent translation.
The initiation of protein synthesis plays an essential regulatory role in human biology. At the center of the initiation pathway, the 13-subunit eukaryotic translation initiation factor 3 (eIF3) controls access of other initiation factors and mRNA to the ribosome by unknown mechanisms. Using electron microscopy (EM), bioinformatics and biochemical experiments, we identify two highly conserved RNA-binding motifs in eIF3 that direct translation initiation from the hepatitis C virus internal ribosome entry site (HCV IRES) RNA. Mutations in the RNA-binding motif of subunit eIF3a weaken eIF3 binding to the HCV IRES and the 40S ribosomal subunit, thereby suppressing eIF2-dependent recognition of the start codon. Mutations in the eIF3c RNA-binding motif also reduce 40S ribosomal subunit binding to eIF3, and inhibit eIF5B-dependent steps downstream of start codon recognition. These results provide the first connection between the structure of the central translation initiation factor eIF3 and recognition of the HCV genomic RNA start codon, molecular interactions that likely extend to the human transcriptome
N-TERMINAL PROCESSING OF RIBOSOMAL PROTEIN L27 IN STAPHYLOCOCCUS AUREUS
The bacterial ribosome is essential to cell growth yet little is known about how its proteins attain their mature structures. Recent studies indicate that certain Staphlyococcus aureus bacteriophage protein sequences contain specific sites that may be cleaved by a non-bacteriophage enzyme (Poliakov et al. 2008). The phage cleavage site was found to bear sequence similarity to the N-terminus of S. aureus ribosomal protein L27. Previous studies in E. coli (Wower et al.1998; Maguire et al. 2005) found that L27 is situated adjacent to the ribosomal peptidyl transferase site, where it likely aids in new peptide formation. The predicted S. aureus L27 protein contains an additional N-terminal sequence not observed within the N-terminus of the otherwise similar E. coli L27; this sequence appears to be cleaved, indicating yet-unobserved ribosomal protein post-translational processing and use of host processes by phage. Phylogenetic analysis shows that L27 processing has the potential to be highly conserved. Further study of this phenomenon may aid antibiotic development
Fungicide Resistance Genetics of Apple Scab fungus Venturia inaequalis
Apple scab, caused by the ascomycete fungus Venturia inaequalis, is considered the most devastating disease on domestic apple crops. Apples are the most important cultivated crop in temperate regions and the United States produced about 4.6 million tons of apples in 2010. Traditional methods to control fungal plant diseases like apple scab are based on the use of chemical compounds that may produce serious negative effects, mainly related with environmental pollution and the development of fungicide resistance. Identifying genes and mechanisms of fungicide resistance in V. inaequalis is imperative to developing new and more effective defenses against the spread of resistance
NASSAM: a server to search for and annotate tertiary interactions and motifs in three-dimensional structures of complex RNA molecules
Similarities in the 3D patterns of RNA base interactions or arrangements can provide insights into their functions and roles in stabilization of the RNA 3D structure. Nucleic Acids Search for Substructures and Motifs (NASSAM) is a graph theoretical program that can search for 3D patterns of base arrangements by representing the bases as pseudo-atoms. The geometric relationship of the pseudo-atoms to each other as a pattern can be represented as a labeled graph where the pseudo-atoms are the graph's nodes while the edges are the inter-pseudo-atomic distances. The input files for NASSAM are PDB formatted 3D coordinates. This web server can be used to identify matches of base arrangement patterns in a query structure to annotated patterns that have been reported in the literature or that have possible functional and structural stabilization implications. The NASSAM program is freely accessible without any login requirement at http://mfrlab.org/grafss/nassam/
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