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      TRPC1 transcript variants, inefficient nonsense-mediated decay and low up-frameshift-1 in vascular smooth muscle cells

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          Abstract

          Background

          Transient Receptor Potential Canonical 1 (TRPC1) is a widely-expressed mammalian cationic channel with functional effects that include stimulation of cardiovascular remodelling. The initial aim of this study was to investigate variation in TRPC1-encoding gene transcripts.

          Results

          Extensive TRPC1 transcript alternative splicing was observed, with exons 2, 3 and 5-9 frequently omitted, leading to variants containing premature termination codons. Consistent with the predicted sensitivity of such variants to nonsense-mediated decay (NMD) the variants were increased by cycloheximide. However it was notable that control of the variants by NMD was prominent in human embryonic kidney 293 cells but not human vascular smooth muscle cells. The cellular difference was attributed in part to a critical protein in NMD, up-frameshift-1 (UPF1), which was found to have low abundance in the vascular cells. Rescue of UPF1 by expression of exogenous UPF1 was found to suppress vascular smooth muscle cell proliferation.

          Conclusions

          The data suggest: (i) extensive NMD-sensitive transcripts of TRPC1; (ii) inefficient clearance of aberrant transcripts and enhanced proliferation of vascular smooth muscle cells in part because of low UPF1 expression.

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          TRP channels.

          Kartik Venkatachalam, Craig Montell (2007)
          The TRP (Transient Receptor Potential) superfamily of cation channels is remarkable in that it displays greater diversity in activation mechanisms and selectivities than any other group of ion channels. The domain organizations of some TRP proteins are also unusual, as they consist of linked channel and enzyme domains. A unifying theme in this group is that TRP proteins play critical roles in sensory physiology, which include contributions to vision, taste, olfaction, hearing, touch, and thermo- and osmosensation. In addition, TRP channels enable individual cells to sense changes in their local environment. Many TRP channels are activated by a variety of different stimuli and function as signal integrators. The TRP superfamily is divided into seven subfamilies: the five group 1 TRPs (TRPC, TRPV, TRPM, TRPN, and TRPA) and two group 2 subfamilies (TRPP and TRPML). TRP channels are important for human health as mutations in at least four TRP channels underlie disease.
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            The nonsense-mediated decay RNA surveillance pathway.

            Yao-Fu Chang, J Saadi Imam, Miles F Wilkinson (2007)
            Nonsense-mediated mRNA decay (NMD) is a quality-control mechanism that selectively degrades mRNAs harboring premature termination (nonsense) codons. If translated, these mRNAs can produce truncated proteins with dominant-negative or deleterious gain-of-function activities. In this review, we describe the molecular mechanism of NMD. We first cover conserved factors known to be involved in NMD in all eukaryotes. We then describe a unique protein complex that is deposited on mammalian mRNAs during splicing, which defines a stop codon as premature. Interaction between this exon-junction complex (EJC) and NMD factors assembled at the upstream stop codon triggers a series of steps that ultimately lead to mRNA decay. We discuss whether these proofreading events preferentially occur during a "pioneer" round of translation in higher and lower eukaryotes, their cellular location, and whether they can use alternative EJC factors or act independent of the EJC.
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              MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments

              Stephen A. Bustin, Jean-François Beaulieu, Jim Huggett … (2010)
              The conclusions of thousands of peer-reviewed publications rely on data obtained using fluorescence-based quantitative real-time PCR technology. However, the inadequate reporting of experimental detail, combined with the frequent use of flawed protocols is leading to the publication of papers that may not be technically appropriate. We take the view that this problem requires the delineation of a more transparent and comprehensive reporting policy from scientific journals. This editorial aims to provide practical guidance for the incorporation of absolute minimum standards encompassing the key assay parameters for accurate design, documentation and reporting of qPCR experiments (MIQE précis) and guidance on the publication of pure 'reference gene' articles.
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                Author and article information

                Journal
                Journal ID (nlm-ta): BMC Mol Biol
                Title: BMC Molecular Biology
                Publisher: BioMed Central
                ISSN (Electronic): 1471-2199
                Publication date Collection: 2011
                Publication date (Electronic): 12 July 2011
                Volume: 12
                Page: 30
                Affiliations
                [1 ]Multidisciplinary Cardiovascular Research Centre
                [2 ]Institute of Membrane & Systems Biology, Faculty of Biological Sciences, Mount Preston Street, University of Leeds, Leeds, LS2 9JT, UK
                [3 ]Yorkshire Heart Centre, General Infirmary at Leeds, Great George Street, Leeds, LS1 3EX, UK
                [4 ]Division of Molecular Immunology, Nikolaus Fiebiger-Center for Molecular Medicine, University of Erlangen-Nürnberg, Glueckstrasse 6, D-91054 Erlangen, Germany
                [5 ]Faculty of Medicine & Health, Clarendon Way, University of Leeds, Leeds, LS2 9JT, UK
                Article
                Publisher ID: 1471-2199-12-30
                DOI: 10.1186/1471-2199-12-30
                PMC ID: 3224110
                PubMed ID: 21749700
                SO-VID: f88909c8-61c9-47f7-9414-306a5cc6d105
                Copyright © Copyright ©2011 Dedman et al; licensee BioMed Central Ltd.
                License:

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Date received : 6 April 2011
                Date accepted : 12 July 2011
                Categories
                Subject: Research Article

                ScienceOpen disciplines: Molecular biology
                Keywords: nonsense-mediated decay,transient receptor potential canonical 1,alternative splicing,cationic channel
                Data availability:
                ScienceOpen disciplines: Molecular biology
                Keywords: nonsense-mediated decay, transient receptor potential canonical 1, alternative splicing, cationic channel

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