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RCDA: a highly sensitive and specific alternatively spliced transcript assembly tool featuring upstream consecutive exon structures.

ABSTRACT: When applied to complex transcript datasets, current tools for automated assembly of mRNA sequences require long run times and produce exponentially increasing numbers of splice variants. Here, we describe RCDA, a genome-based transcript assembly tool comprising RCluster, that recursively clusters transcripts, and DAssemble, that generates composite transcript sequences through path-finding using a directed acyclic graph. Each exon included in a final transcript is associated with an array of all upstream consecutive exon structures obtained from original transcripts. When a depth-first-search path reaches an exon, the path is retained only if it contains a structure from that exon's array. RCDA assemblies, therefore, include only those transcripts with experimentally supported exon patterns. When applied to >23,000 transcripts from human chromosome 21, using biologically reasonable filters, RCDA execution time was approximately 4h. RCDA outperformed ECgene in reconstructing RefSeq transcripts and in limiting the total number of transcripts and transcripts per gene.

SUBMITTER: Sturgeon XH

PROVIDER: S-EPMC5470730 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature

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RCDA: a highly sensitive and specific alternatively spliced transcript assembly tool featuring upstream consecutive exon structures.

Sturgeon Xiaolu H XH Gardiner Katheleen J KJ

Genomics 20120820 6

When applied to complex transcript datasets, current tools for automated assembly of mRNA sequences require long run times and produce exponentially increasing numbers of splice variants. Here, we describe RCDA, a genome-based transcript assembly tool comprising RCluster, that recursively clusters transcripts, and DAssemble, that generates composite transcript sequences through path-finding using a directed acyclic graph. Each exon included in a final transcript is associated with an array of al ...[more]

PMID: 22971325

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Project description:We have examined the regulation of mutually exclusive Cav2.2 exon 37a and b variants by the mouse ?-opioid receptor (mMOR) C-terminal splice variants 1, 1C and 1O in tsA-201 cells. Electrophysiological analyses revealed that both channel isoforms exhibit DAMGO-induced voltage-dependent (G??-mediated) inhibition and its recovery by voltage pre-pulses, as well as a voltage-independent component. However, the two channel isoforms differ in their relative extent of voltage-dependent and independent inhibition, with Cav2.2-37b showing significantly more voltage-dependent inhibition upon activation of the three mMOR receptors studied. In addition, coexpression of either mMOR1 or mMOR1C results in an agonist-independent reduction in the peak current density of Cav2.2-37a channels, whereas the peak current density of Cav2.2-37b does not appear to be affected. Interestingly, this decrease is not due to an effect on channel expression at the plasma membrane, as demonstrated by biotinylation experiments. We further examined the mechanism underlying the agonist-independent modulation of Cav2.2-37a by mMOR1C. Incubation of cells with pertussis toxin did not affect the mMOR1C mediated inhibition of Cav2.2-37a currents, indicating a lack of involvement of Gi/o signaling. However, when a Src tyrosine kinase inhibitor was applied, the effect of mMOR1C was lost. Moreover, when we recorded currents using a Cav2.2-37a mutant in which tyrosine 1747 was replaced with phenylalanine (Y1747F), the agonist independent effects of mMOR1C were abolished. Altogether our findings show that Cav2.2-37a and Cav2.2-37b isoforms are subject to differential regulation by C-terminal splice variants of mMORs, and that constitutive mMOR1C activity and downstream tyrosine kinase activity exert a selective inhibition of the Cav2.2-37a splice variant, an N-type channel isoform that is highly enriched in nociceptors. Our study provides new insights into the roles of the MOR full-length C-terminal variants in modulating Cav2.2 channel isoform activities.

Dataset Information

RCDA: a highly sensitive and specific alternatively spliced transcript assembly tool featuring upstream consecutive exon structures.

Publications

RCDA: a highly sensitive and specific alternatively spliced transcript assembly tool featuring upstream consecutive exon structures.

Similar Datasets

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