RNA CUG Repeats Sequester CUGBP1 and Alter Protein Levels and Activity of CUGBP1

Timchenko, Nikolai A.; Cai, Zong Jin; Welm, Alana L.; Reddy, Sita; Ashizawa, Tetsuo; Timchenko, Lubov

doi:10.1074/jbc.m005960200

Cited by 273 publications

(290 citation statements)

References 29 publications

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“…CUGBP1 was detected with polyclonal antibodies against CUGBP1 (1:1000) as described. 14 To determine protein loading, the same membrane after CUGBP1 detection was reprobed with antibodies against b-actin, and CUGBP1 levels were calculated as ratio to b-actin.…”

Section: Western Analysismentioning

confidence: 99%

“…[2][3][4] At the molecular levels, the DM1 phenotype is most likely caused by a complex molecular pathogenesis, including deficiency of myotonic dystrophy myotonin kinase (DMPK) protein, [5][6][7] haplo-insufficiency of a neighboring homeobox gene (particular the DM locus-associated homeodomain protein (DMAHP/Six 5 gene) 8 and the WD-repeat gene (DMWD) 9 and a trans-dominant misregulation of RNA homeostasis. [10][11][12][13][14][15][16] Recent experiments from transgenic mice, expressing an untranslated expanded CUG repeat under the control of the human skeletal actin promoter, showed that expanded CUG repeats are sufficient to generate DM1 muscle phenotype. 17 These data suggest that misregulation of RNA homeostasis may play a major role in DM1 muscle pathogenesis.…”

Section: Introductionmentioning

confidence: 99%

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Viral vector producing antisense RNA restores myotonic dystrophy myoblast functions

Furling

Doucet²,

Ma³

et al. 2003

Gene Ther

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Myotonic dystrophy (DM1) is caused by the expansion of a trinucleotide repeat (CTG) located in the 3 0 untranslated region of the myotonic dystrophy protein kinase gene, for which currently there is no effective treatment. The data available suggest that misregulation of RNA homeostasis may play a major role in DM1 muscle pathogenesis. This indicates that the specific targeting of the mutant DMPK transcripts is essential to raise the rationale basis for the development of a specific gene therapy for DM1. We have produced a retrovirus which expresses a 149-bp antisense RNA complementary to the (CUG)13 repeats and to the 110-bp region following the repeats sequence to increase the specificity. This construct was introduced into human DM1 myoblasts, resulting in a preferential decrease in mutant DMPK transcripts, and effective restoration of human DM1 myoblast functions such as myoblast fusion and the uptake of glucose. It was previously shown that delay of muscle differentiation and insulin resistance in DM1 are associated with misregulation of CUGBP1 protein levels. The analysis of CUGBP1 levels and activity in DM1 cells expressing the antisense RNA indicated a correction of CUGBP1 expression in infected DM1 cells. We therefore show that current antisense RNA delivered in vitro using a retrovirus is not only capable of inhibiting mutant DMPK transcripts, but also can ameliorate dystrophic muscle pathology at the cellular levels. Gene Therapy (2003) 10, 795-802.

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Section: Western Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Viral vector producing antisense RNA restores myotonic dystrophy myoblast functions

Furling

Doucet²,

Ma³

et al. 2003

Gene Ther

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“…One of the members of the CUGBP family, CUGBP1, contributes to at least three symptoms of DM1: the delay of skeletal muscle differentiation (12), myotonia (19), and the insulin resistance (13). Previous studies showed that the expansion of CUG repeats in DM1 leads to a 3-5-fold elevation of CUGBP1 binding activity and protein levels (13, 19 -21), perhaps by stabilizing CUGBP1 protein within RNA CUG-CUGBP1 complexes (21). CUGBP1 possesses two major biological functions: regulation of splicing and translation (12, 20 -22).…”

Section: Dm1mentioning

confidence: 99%

Overexpression of CUG Triplet Repeat-binding Protein, CUGBP1, in Mice Inhibits Myogenesis

Timchenko

Patel

Iakova

et al. 2004

Journal of Biological Chemistry

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205

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Accumulation of RNA CUG repeats in myotonic dystrophy type 1 (DM1) patients leads to the induction of a CUG-binding protein, CUGBP1, which increases translation of several proteins that are required for myogenesis. In this paper, we examine the role of overexpression of CUGBP1 in DM1 muscle pathology using transgenic mice that overexpress CUGBP1 in skeletal muscle. Our data demonstrate that the elevation of CUGBP1 in skeletal muscle causes overexpression of MEF2A and p21 to levels that are significantly higher than those in skeletal muscle of wild type animals. A similar induction of these proteins is observed in skeletal muscle of DM1 patients with increased levels of CUGBP1. Immunohistological analysis showed that the skeletal muscle from mice overexpressing CUGBP1 is characterized by a developmental delay, muscular dystrophy, and myofiber-type switch: increase of slow/oxidative fibers and the reduction of fast fibers. Examination of molecular mechanisms by which CUGBP1 up-regulates MEF2A shows that CUGBP1 increases translation of MEF2A via direct interaction with GCN repeats located within MEF2A mRNA. Our data suggest that CUGBP1-mediated overexpression of MEF2A and p21 inhibits myogenesis and contributes to the development of muscle deficiency in DM1 patients. DM11 is a multisystem disease mainly characterized by defects in skeletal muscle with the involvement of many tissues and systems such as cardiac muscle, brain, eye, and endocrine system (1). DM1 is caused by an expansion of CTG trinucleotide repeats within the 3Ј-untranslated region of the myotonin protein kinase gene (2). In DM1 patients, the size of DNA CTG expansion correlates with the severity of the disease. Patients with CTG expansion containing 50 -80 CTG repeats are almost asymptomatic. Individuals bearing the myotonin protein kinase gene with 100 -500 CTG repeats develop a disease in adult life (classical adult form of DM1) that is characterized by a progressive muscle wasting with myotonia. The most severe form of DM1, congenital disease, affects patients before or after birth and is associated with long CTG expansions (up to 2,000 repeats). This form of disease is characterized by a delay or arrest of skeletal muscle development (3). Although there is an overlap in range of repeats between different forms of DM1, there is a clear correlation of repeat number with severity of phenotype and reduction of age of onset.Investigations of molecular alterations in DM1 suggest that the expansion of CTG repeats causes the DM1 pathology through different mechanisms, mediated at both DNA and RNA levels. It has been shown that CTG repeats reduce expression of myotonin protein kinase in cis (4), causing abnormalities in cardiac muscle (5). CTG repeats also affect transcription of genes adjacent to myotonin protein kinase (6), leading to the development of cataracts (7,8). A number of recent studies indicate that other symptoms in DM1 such as myotonia (9, 10), delay of skeletal muscle differentiation (11,12), and a resistance to insulin (13) are mediated ...

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“…The main problem of this strategy is low stability of siRNA and lack of efficient delivery systems of these reagents. 12,13 The complex proposed in our study should effectively bind and stabilise siRNA for transfection and should show low toxicity for a variety of cells. For the lactose based surfactants chosen (LA12), the bonding of lactose moiety with the hydrophobic fragment via an amine group ensures effective proton bonding in acidic environment, which means that they can act as cationic surfactants.…”

mentioning

confidence: 99%

“…The sequence studied is responsible for silencing the gene that can be important in therapy of myotonic dystrophy type 1 (DM1). [11][12][13] DM1 is the most frequent form of muscular dystrophy in adults. 14 It is an autosomal-dominant hereditary disease which can affect many tissues and produce diverse symptoms, including muscle hyperactivity (myotonia), progressive myasthenia, heart conduction defects, cardiomyopathy, insulin resistance, or neuropsychological disturbances.…”

mentioning

confidence: 99%

The system with zwitterionic lactose-based surfactant for complexation and delivery of small interfering ribonucleic acid—A structural and spectroscopic study

Skupin

Sobczak

Zieliński

et al. 2016

Applied Physics Letters

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Systems suitable for the effective preparation of complexes with siRNA (small interfering RNA) are at the center of interest in the area of research work on the delivery of the RNA-based drugs (RNA-therapeutics). This article presents results of a study on the structural effects associated with siRNA complexation by a surfactant comprising a lactose group (N-(3-propanesulfone)-N-dodecyl-amino-beta-D-lactose hydrochloride, LA12). The double stranded siRNA oligomer (21 base pairs) used in this study is responsible for silencing a gene that can be important in the therapy of myotonic dystrophy type 1. The obtained siRNA/LA12 lipoplexes were studied using the methods of small angle scattering of synchrotron radiation, circular dichroism spectroscopy, Fourier transform infrared spectroscopy, and electrophoretic mobility tests. Lipoplexes form in solution stable lamellar or cubic phases. The surfactant selected for the study shows much lower cytotoxicity and good complexation abilities of siRNA than dicationic or polycationic surfactants.

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RNA CUG Repeats Sequester CUGBP1 and Alter Protein Levels and Activity of CUGBP1

Cited by 273 publications

References 29 publications

Viral vector producing antisense RNA restores myotonic dystrophy myoblast functions

Viral vector producing antisense RNA restores myotonic dystrophy myoblast functions

Overexpression of CUG Triplet Repeat-binding Protein, CUGBP1, in Mice Inhibits Myogenesis

The system with zwitterionic lactose-based surfactant for complexation and delivery of small interfering ribonucleic acid—A structural and spectroscopic study

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