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  1. Article ; Online: Riboswitches as hormone receptors: hypothetical cytokinin-binding riboswitches in Arabidopsis thaliana.

    Grojean, Jeremy / Downes, Brian

    Biology direct

    2010  Volume 5, Page(s) 60

    Abstract: Background: Riboswitches are mRNA elements that change conformation when bound to small molecules. They are known to be key regulators of biosynthetic pathways in both prokaryotes and eukaryotes.: Presentation of the hypothesis: The hypothesis ... ...

    Abstract Background: Riboswitches are mRNA elements that change conformation when bound to small molecules. They are known to be key regulators of biosynthetic pathways in both prokaryotes and eukaryotes.
    Presentation of the hypothesis: The hypothesis presented here is that riboswitches function as receptors in hormone perception. We propose that riboswitches initiate or integrate signaling cascades upon binding to classic signaling molecules. The molecular interactions for ligand binding and gene expression control would be the same as for biosynthetic pathways, but the context and the cadre of ligands to consider is dramatically different. The hypothesis arose from the observation that a compound used to identify adenine binding RNA sequences is chemically similar to the classic plant hormone, or growth regulator, cytokinin. A general tenet of the hypothesis is that riboswitch-binding metabolites can be used to make predictions about chemically related signaling molecules. In fact, all cell permeable signaling compounds can be considered as potential riboswitch ligands. The hypothesis is plausible, as demonstrated by a cursory review of the transcriptome and genome of the model plant Arabidopsis thaliana for transcripts that i) contain an adenine aptamer motif, and ii) are also predicted to be cytokinin-regulated. Here, one gene, CRK10 (for Cysteine-rich Receptor-like Kinase 10, At4g23180), contains an adenine aptamer-related sequence and is down-regulated by cytokinin approximately three-fold in public gene expression data. To illustrate the hypothesis, implications of cytokinin-binding to the CRK10 mRNA are discussed.
    Testing the hypothesis: At the broadest level, screening various cell permeable signaling molecules against random RNA libraries and comparing hits to sequence and gene expression data bases could determine how broadly the hypothesis applies. Specific cases, such as CRK10 presented here, will require experimental validation of direct ligand binding, altered RNA conformation, and effect on gene expression. Each case will be different depending on the signaling pathway and the physiology involved.
    Implications of the hypothesis: This would be a very direct signal perception mechanism for regulating gene expression; rivaling animal steroid hormone receptors, which are frequently ligand dependent transcription initiation factors. Riboswitch-regulated responses could occur by modulating target RNA stability, translatability, and alternative splicing - all known expression platforms used in riboswitches. The specific illustration presented, CRK10, implies a new mechanism for the perception of cytokinin, a classic plant hormone. Experimental support for the hypothesis would add breadth to the growing list of important functions attributed to riboswitches.
    Reviewers: This article was reviewed by Anthony Poole, Rob Knight, Mikhail Gelfand.
    MeSH term(s) Arabidopsis/genetics ; Arabidopsis/metabolism ; Cytokinins/metabolism ; RNA, Plant/genetics ; RNA, Plant/metabolism ; Riboswitch/genetics ; Riboswitch/physiology
    Chemical Substances Cytokinins ; RNA, Plant ; Riboswitch
    Language English
    Publishing date 2010-10-20
    Publishing country England
    Document type Journal Article
    ISSN 1745-6150
    ISSN (online) 1745-6150
    DOI 10.1186/1745-6150-5-60
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Rchimerism: An R Package for Automated Chimerism Data Analysis.

    Siddiqui, Zohair / Maldonado, Juan / Grojean, Jeremy / Ye, Fei / Zhang, David / Longtine, Janina / Ahn, Tae-Hyuk / Guo, Huazhang

    The Journal of molecular diagnostics : JMD

    2019  Volume 22, Issue 1, Page(s) 21–29

    Abstract: A quantitative chimerism test monitors engraftment of donor hematopoietic stem cells or relapse of leukemias or lymphomas in hematopoietic stem cell transplantation patients. The most common method used for chimerism testing is PCR amplification of short ...

    Abstract A quantitative chimerism test monitors engraftment of donor hematopoietic stem cells or relapse of leukemias or lymphomas in hematopoietic stem cell transplantation patients. The most common method used for chimerism testing is PCR amplification of short tandem repeat loci, followed by capillary gel electrophoresis. Manual data analysis is tedious and time consuming, as it involves the selection of informative loci and the repetition of quantifying chimerism percentage for multiple loci from multiple cell types. It is also susceptible to human errors. Currently, there is no free software to fully automate chimerism data analysis. Rchimerism, an R shiny package, was developed to automatically pick informative loci, calculate chimerism percentage, and display the results through a user-friendly interface. The accuracy of the program was compared with manual calculation on 60 patient samples with 100% concordance. Compared with manual calculation, Rchimerism drastically reduces analysis time from 20 to 40 minutes for single donor transplantation samples and from 40 to 80 minutes for double donor transplantation samples to >1 minute. Rchimerism can be downloaded and used freely by noncommercial laboratories.
    MeSH term(s) Algorithms ; Alleles ; Biomarkers ; Chimera/genetics ; Chimerism ; Data Accuracy ; Data Analysis ; Electrophoresis, Capillary ; Genetic Loci ; Graft Rejection/genetics ; Graft Survival/genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Microsatellite Repeats ; Polymerase Chain Reaction ; Tissue Donors ; Transplant Recipients ; User-Computer Interface
    Chemical Substances Biomarkers
    Language English
    Publishing date 2019-10-09
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Validation Study
    ZDB-ID 2000060-1
    ISSN 1943-7811 ; 1525-1578
    ISSN (online) 1943-7811
    ISSN 1525-1578
    DOI 10.1016/j.jmoldx.2019.08.005
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Riboswitches as hormone receptors

    Downes Brian / Grojean Jeremy

    Biology Direct, Vol 5, Iss 1, p

    hypothetical cytokinin-binding riboswitches in Arabidopsis thaliana

    2010  Volume 60

    Abstract: Abstract Background Riboswitches are mRNA elements that change conformation when bound to small molecules. They are known to be key regulators of biosynthetic pathways in both prokaryotes and eukaryotes. Presentation of the Hypothesis The hypothesis ... ...

    Abstract Abstract Background Riboswitches are mRNA elements that change conformation when bound to small molecules. They are known to be key regulators of biosynthetic pathways in both prokaryotes and eukaryotes. Presentation of the Hypothesis The hypothesis presented here is that riboswitches function as receptors in hormone perception. We propose that riboswitches initiate or integrate signaling cascades upon binding to classic signaling molecules. The molecular interactions for ligand binding and gene expression control would be the same as for biosynthetic pathways, but the context and the cadre of ligands to consider is dramatically different. The hypothesis arose from the observation that a compound used to identify adenine binding RNA sequences is chemically similar to the classic plant hormone, or growth regulator, cytokinin. A general tenet of the hypothesis is that riboswitch-binding metabolites can be used to make predictions about chemically related signaling molecules. In fact, all cell permeable signaling compounds can be considered as potential riboswitch ligands. The hypothesis is plausible, as demonstrated by a cursory review of the transcriptome and genome of the model plant Arabidopsis thaliana for transcripts that i) contain an adenine aptamer motif, and ii) are also predicted to be cytokinin-regulated. Here, one gene, CRK10 (for Cysteine-rich Receptor-like Kinase 10 , At4g23180), contains an adenine aptamer-related sequence and is down-regulated by cytokinin approximately three-fold in public gene expression data. To illustrate the hypothesis, implications of cytokinin-binding to the CRK10 mRNA are discussed. Testing the hypothesis At the broadest level, screening various cell permeable signaling molecules against random RNA libraries and comparing hits to sequence and gene expression data bases could determine how broadly the hypothesis applies. Specific cases, such as CRK10 presented here, will require experimental validation of direct ligand binding, altered RNA conformation, and effect on gene ...
    Keywords Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2010-10-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article: Rho-kinase-mediated Ca2+-independent contraction in rat embryo fibroblasts.

    Emmert, Daniel A / Fee, Judy A / Goeckeler, Zoe M / Grojean, Jeremy M / Wakatsuki, Tetsuro / Elson, Elliot L / Herring, B Paul / Gallagher, Patricia J / Wysolmerski, Robert B

    American journal of physiology. Cell physiology

    2003  Volume 286, Issue 1, Page(s) C8–21

    Abstract: Thus far, determining the relative contribution of Ca2+/calmodulin-dependent myosin light chain kinase (MLCK) and Ca2+-independent Rho-kinase pathways to myosin II activation and contraction has been difficult. In this study, we characterize the role of ... ...

    Abstract Thus far, determining the relative contribution of Ca2+/calmodulin-dependent myosin light chain kinase (MLCK) and Ca2+-independent Rho-kinase pathways to myosin II activation and contraction has been difficult. In this study, we characterize the role of Rho-kinase in a rat embryo fibroblast cell line (REF-52), which contains no detectable MLCK. No endogenous MLCK could be detected in REF-52 cells by either Western or Northern blot analysis. In the presence or absence of Ca2+, thrombin or lysophosphatidic acid (LPA) increased RhoA activity and Rhokinase activity, correlating with isometric tension development and myosin II regulatory light chain (RLC) phosphorylation. Resting tension is associated with a basal phosphorylation of 0.31 +/- 0.02 mol PO4/mol RLC, whereas upon LPA or thrombin treatment myosin II RLC phosphorylation increases to 1.08 +/- 0.05 and 0.82 +/- 0.05 mol PO4/mol RLC, respectively, within 2.5 min. Ca2+ chelation has minimal effect on the kinetics and magnitude of isometric tension development and RLC phosphorylation. Treatment of REF-52 cells with the Rho-kinase-specific inhibitor Y-27632 abolished thrombin- and LPA-stimulated contraction and RLC phosphorylation. These results suggest that Rho-kinase is sufficient to activate myosin II motor activity and contraction in REF-52 cells.
    MeSH term(s) Amides/pharmacology ; Animals ; Calcium/physiology ; Calcium-Calmodulin-Dependent Protein Kinases/metabolism ; Cell Line ; Embryo, Mammalian ; Enzyme Inhibitors/pharmacology ; Fibroblasts/drug effects ; Fibroblasts/physiology ; Intracellular Signaling Peptides and Proteins ; Lysophospholipids/pharmacology ; Myosin Light Chains/metabolism ; Myosin-Light-Chain Kinase/metabolism ; Phosphorylation ; Protein-Serine-Threonine Kinases/metabolism ; Protein-Serine-Threonine Kinases/physiology ; Pyridines/pharmacology ; Rats ; Thrombin/pharmacology ; rho-Associated Kinases ; rhoA GTP-Binding Protein/metabolism
    Chemical Substances Amides ; Enzyme Inhibitors ; Intracellular Signaling Peptides and Proteins ; Lysophospholipids ; Myosin Light Chains ; Pyridines ; Y 27632 (138381-45-0) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; rho-Associated Kinases (EC 2.7.11.1) ; Calcium-Calmodulin-Dependent Protein Kinases (EC 2.7.11.17) ; Myosin-Light-Chain Kinase (EC 2.7.11.18) ; Thrombin (EC 3.4.21.5) ; rhoA GTP-Binding Protein (EC 3.6.5.2) ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2003-09-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 392098-7
    ISSN 1522-1563 ; 0363-6143
    ISSN (online) 1522-1563
    ISSN 0363-6143
    DOI 10.1152/ajpcell.00428.2002
    Database MEDical Literature Analysis and Retrieval System OnLINE

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