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  1. Article: Identification of over ten thousand candidate structured RNAs in viruses and phages.

    Fremin, Brayon J / Bhatt, Ami S / Kyrpides, Nikos C

    Computational and structural biotechnology journal

    2023  Volume 21, Page(s) 5630–5639

    Abstract: Structured RNAs play crucial roles in viruses, exerting influence over both viral and host gene expression. However, the extensive diversity of structured RNAs and their ability to act in cis or trans positions pose challenges for predicting and ... ...

    Abstract Structured RNAs play crucial roles in viruses, exerting influence over both viral and host gene expression. However, the extensive diversity of structured RNAs and their ability to act in cis or trans positions pose challenges for predicting and assigning their functions. While comparative genomics approaches have successfully predicted candidate structured RNAs in microbes on a large scale, similar efforts for viruses have been lacking. In this study, we screened over 5 million DNA and RNA viral sequences, resulting in the prediction of 10,006 novel candidate structured RNAs. These predictions are widely distributed across taxonomy and ecosystem. We found transcriptional evidence for 206 of these candidate structured RNAs in the human fecal microbiome. These candidate RNAs exhibited evidence of nucleotide covariation, indicative of selective pressure maintaining the predicted secondary structures. Our analysis revealed a diverse repertoire of candidate structured RNAs, encompassing a substantial number of putative tRNAs or tRNA-like structures, Rho-independent transcription terminators, and potentially cis-regulatory structures consistently positioned upstream of genes. In summary, our findings shed light on the extensive diversity of structured RNAs in viruses, offering a valuable resource for further investigations into their functional roles and implications in viral gene expression and pave the way for a deeper understanding of the intricate interplay between viruses and their hosts at the molecular level.
    Language English
    Publishing date 2023-11-07
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2694435-2
    ISSN 2001-0370
    ISSN 2001-0370
    DOI 10.1016/j.csbj.2023.11.010
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Identifying candidate structured RNAs in CRISPR operons.

    Fremin, Brayon J / Kyrpides, Nikos C

    RNA biology

    2021  Volume 19, Issue 1, Page(s) 678–685

    Abstract: Noncoding RNAs with secondary structures play important roles in CRISPR-Cas systems. Many of these structures likely remain undiscovered. We used a large-scale comparative genomics approach to predict 156 novel candidate structured RNAs from 36,111 ... ...

    Abstract Noncoding RNAs with secondary structures play important roles in CRISPR-Cas systems. Many of these structures likely remain undiscovered. We used a large-scale comparative genomics approach to predict 156 novel candidate structured RNAs from 36,111 CRISPR-Cas systems. A number of these were found to overlap with coding genes, including palindromic candidates that overlapped with a variety of Cas genes in type I and III systems. Among these 156 candidates, we identified 46 new models of CRISPR direct repeats and 1 tracrRNA. This tracrRNA model occasionally overlapped with predicted
    MeSH term(s) CRISPR-Cas Systems ; Genomics ; Operon ; RNA/genetics ; Repetitive Sequences, Nucleic Acid
    Chemical Substances RNA (63231-63-0)
    Language English
    Publishing date 2021-12-31
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2159587-2
    ISSN 1555-8584 ; 1555-8584
    ISSN (online) 1555-8584
    ISSN 1555-8584
    DOI 10.1080/15476286.2022.2067714
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Comparative genomics identifies thousands of candidate structured RNAs in human microbiomes.

    Fremin, Brayon J / Bhatt, Ami S

    Genome biology

    2021  Volume 22, Issue 1, Page(s) 100

    Abstract: Background: Structured RNAs play varied bioregulatory roles within microbes. To date, hundreds of candidate structured RNAs have been predicted using informatic approaches that search for motif structures in genomic sequence data. The human microbiome ... ...

    Abstract Background: Structured RNAs play varied bioregulatory roles within microbes. To date, hundreds of candidate structured RNAs have been predicted using informatic approaches that search for motif structures in genomic sequence data. The human microbiome contains thousands of species and strains of microbes. Yet, much of the metagenomic data from the human microbiome remains unmined for structured RNA motifs primarily due to computational limitations.
    Results: We sought to apply a large-scale, comparative genomics approach to these organisms to identify candidate structured RNAs. With a carefully constructed, though computationally intensive automated analysis, we identify 3161 conserved candidate structured RNAs in intergenic regions, as well as 2022 additional candidate structured RNAs that may overlap coding regions. We validate the RNA expression of 177 of these candidate structures by analyzing small fragment RNA-seq data from four human fecal samples.
    Conclusions: This approach identifies a wide variety of candidate structured RNAs, including tmRNAs, antitoxins, and likely ribosome protein leaders, from a wide variety of taxa. Overall, our pipeline enables conservative predictions of thousands of novel candidate structured RNAs from human microbiomes.
    MeSH term(s) Computational Biology/methods ; Gastrointestinal Microbiome ; Genomics/methods ; Humans ; Metagenome ; Metagenomics/methods ; Microbiota ; Nucleic Acid Conformation ; RNA ; Workflow
    Chemical Substances RNA (63231-63-0)
    Language English
    Publishing date 2021-04-12
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2040529-7
    ISSN 1474-760X ; 1474-760X
    ISSN (online) 1474-760X
    ISSN 1474-760X
    DOI 10.1186/s13059-021-02319-w
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Structured RNA Contaminants in Bacterial Ribo-Seq.

    Fremin, Brayon J / Bhatt, Ami S

    mSphere

    2020  Volume 5, Issue 5

    Abstract: Ribosome profiling (Ribo-Seq) is a powerful method to study translation in bacteria. However, Ribo-Seq signal can be observed across RNAs that one would not expect to be bound by ribosomes. For example, ...

    Abstract Ribosome profiling (Ribo-Seq) is a powerful method to study translation in bacteria. However, Ribo-Seq signal can be observed across RNAs that one would not expect to be bound by ribosomes. For example,
    MeSH term(s) Computational Biology ; Escherichia coli/genetics ; Gene Expression Profiling ; RNA, Untranslated/chemistry ; RNA, Untranslated/genetics ; Ribosomes/genetics ; Sequence Analysis, RNA
    Chemical Substances RNA, Untranslated
    Language English
    Publishing date 2020-10-21
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ISSN 2379-5042
    ISSN (online) 2379-5042
    DOI 10.1128/mSphere.00855-20
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Thousands of small, novel genes predicted in global phage genomes.

    Fremin, Brayon J / Bhatt, Ami S / Kyrpides, Nikos C

    Cell reports

    2022  Volume 39, Issue 12, Page(s) 110984

    Abstract: Small genes ( ... 40,000 small-gene families in ∼2.3 million phage genome contigs. We find that small genes in phage ... ...

    Abstract Small genes (<150 nucleotides) have been systematically overlooked in phage genomes. We employ a large-scale comparative genomics approach to predict >40,000 small-gene families in ∼2.3 million phage genome contigs. We find that small genes in phage genomes are approximately 3-fold more prevalent than in host prokaryotic genomes. Our approach enriches for small genes that are translated in microbiomes, suggesting the small genes identified are coding. More than 9,000 families encode potentially secreted or transmembrane proteins, more than 5,000 families encode predicted anti-CRISPR proteins, and more than 500 families encode predicted antimicrobial proteins. By combining homology and genomic-neighborhood analyses, we reveal substantial novelty and diversity within phage biology, including small phage genes found in multiple host phyla, small genes encoding proteins that play essential roles in host infection, and small genes that share genomic neighborhoods and whose encoded proteins may share related functions.
    MeSH term(s) Bacteriophages/genetics ; Genome, Viral/genetics ; Genomics ; Microbiota ; Phylogeny
    Language English
    Publishing date 2022-06-23
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2022.110984
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Identifying candidate structured RNAs in CRISPR operons

    Fremin, Brayon J. / Kyrpides, Nikos C.

    RNA Biology. 2022 Dec. 31, v. 19, no. 1 p.678-685

    2022  

    Abstract: Noncoding RNAs with secondary structures play important roles in CRISPR-Cas systems. Many of these structures likely remain undiscovered. We used a large-scale comparative genomics approach to predict 156 novel candidate structured RNAs from 36,111 ... ...

    Abstract Noncoding RNAs with secondary structures play important roles in CRISPR-Cas systems. Many of these structures likely remain undiscovered. We used a large-scale comparative genomics approach to predict 156 novel candidate structured RNAs from 36,111 CRISPR-Cas systems. A number of these were found to overlap with coding genes, including palindromic candidates that overlapped with a variety of Cas genes in type I and III systems. Among these 156 candidates, we identified 46 new models of CRISPR direct repeats and 1 tracrRNA. This tracrRNA model occasionally overlapped with predicted cas9 coding regions, emphasizing the importance of expanding our search windows for novel structure RNAs in coding regions. We also demonstrated that the antirepeat sequence in this tracrRNA model can be used to accurately assign thousands of predicted CRISPR arrays to type II-C systems. This study highlights the importance of unbiased identification of candidate structured RNAs across CRISPR-Cas systems.
    Keywords CRISPR-Cas systems ; RNA ; genomics ; models ; operon ; CRISPR ; structured RNA ; comparative genomics
    Language English
    Dates of publication 2022-1231
    Size p. 678-685.
    Publishing place Taylor & Francis
    Document type Article ; Online
    ZDB-ID 2159587-2
    ISSN 1555-8584
    ISSN 1555-8584
    DOI 10.1080/15476286.2022.2067714
    Database NAL-Catalogue (AGRICOLA)

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  7. Article ; Online: Simultaneous ribosome profiling of hundreds of microbes from the human microbiome.

    Fremin, Brayon J / Nicolaou, Cosmos / Bhatt, Ami S

    Nature protocols

    2021  Volume 16, Issue 10, Page(s) 4676–4691

    Abstract: Ribosome profiling enables sequencing of ribosome-bound fragments of RNA, revealing which transcripts are being translated as well as the position of ribosomes along mRNAs. Although ribosome profiling has been applied to cultured bacterial isolates, its ... ...

    Abstract Ribosome profiling enables sequencing of ribosome-bound fragments of RNA, revealing which transcripts are being translated as well as the position of ribosomes along mRNAs. Although ribosome profiling has been applied to cultured bacterial isolates, its application to uncultured, mixed communities has been challenging. We present MetaRibo-Seq, a protocol that enables the application of ribosome profiling directly to the human fecal microbiome. MetaRibo-Seq is a benchmarked method that includes several modifications to existing ribosome profiling protocols, specifically addressing challenges involving fecal sample storage, purity and input requirements. We also provide a computational workflow to quality control and trim reads, de novo assemble a reference metagenome with metagenomic reads, align MetaRibo-Seq reads to the reference, and assess MetaRibo-Seq library quality ( https://github.com/bhattlab/bhattlab_workflows/tree/master/metariboseq ). This MetaRibo-Seq protocol enables researchers in standard molecular biology laboratories to study translation in the fecal microbiome in ~5 d.
    MeSH term(s) Feces ; Humans ; Microbiota ; Ribosomes ; Sequence Analysis, RNA
    Language English
    Publishing date 2021-08-11
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2244966-8
    ISSN 1750-2799 ; 1754-2189
    ISSN (online) 1750-2799
    ISSN 1754-2189
    DOI 10.1038/s41596-021-00592-4
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Comparative genomics identifies thousands of candidate structured RNAs in human microbiomes

    Brayon J. Fremin / Ami S. Bhatt

    Genome Biology, Vol 22, Iss 1, Pp 1-

    2021  Volume 16

    Abstract: Abstract Background Structured RNAs play varied bioregulatory roles within microbes. To date, hundreds of candidate structured RNAs have been predicted using informatic approaches that search for motif structures in genomic sequence data. The human ... ...

    Abstract Abstract Background Structured RNAs play varied bioregulatory roles within microbes. To date, hundreds of candidate structured RNAs have been predicted using informatic approaches that search for motif structures in genomic sequence data. The human microbiome contains thousands of species and strains of microbes. Yet, much of the metagenomic data from the human microbiome remains unmined for structured RNA motifs primarily due to computational limitations. Results We sought to apply a large-scale, comparative genomics approach to these organisms to identify candidate structured RNAs. With a carefully constructed, though computationally intensive automated analysis, we identify 3161 conserved candidate structured RNAs in intergenic regions, as well as 2022 additional candidate structured RNAs that may overlap coding regions. We validate the RNA expression of 177 of these candidate structures by analyzing small fragment RNA-seq data from four human fecal samples. Conclusions This approach identifies a wide variety of candidate structured RNAs, including tmRNAs, antitoxins, and likely ribosome protein leaders, from a wide variety of taxa. Overall, our pipeline enables conservative predictions of thousands of novel candidate structured RNAs from human microbiomes.
    Keywords Biology (General) ; QH301-705.5 ; Genetics ; QH426-470
    Subject code 500
    Language English
    Publishing date 2021-04-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: MetaRibo-Seq measures translation in microbiomes

    Brayon J. Fremin / Hila Sberro / Ami S. Bhatt

    Nature Communications, Vol 11, Iss 1, Pp 1-

    2020  Volume 12

    Abstract: Defining the functions of individual organisms or communities within microbiomes is a challenging task. Here, the authors develop MetaRibo-Seq, a method for simultaneous high-throughput ribosome profiling of organisms in uncultured microbiome samples. ...

    Abstract Defining the functions of individual organisms or communities within microbiomes is a challenging task. Here, the authors develop MetaRibo-Seq, a method for simultaneous high-throughput ribosome profiling of organisms in uncultured microbiome samples.
    Keywords Science ; Q
    Language English
    Publishing date 2020-06-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: MetaRibo-Seq measures translation in microbiomes

    Brayon J. Fremin / Hila Sberro / Ami S. Bhatt

    Nature Communications, Vol 11, Iss 1, Pp 1-

    2020  Volume 12

    Abstract: Defining the functions of individual organisms or communities within microbiomes is a challenging task. Here, the authors develop MetaRibo-Seq, a method for simultaneous high-throughput ribosome profiling of organisms in uncultured microbiome samples. ...

    Abstract Defining the functions of individual organisms or communities within microbiomes is a challenging task. Here, the authors develop MetaRibo-Seq, a method for simultaneous high-throughput ribosome profiling of organisms in uncultured microbiome samples.
    Keywords Science ; Q
    Language English
    Publishing date 2020-06-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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