title: In vivo co-localisation of MBNL protein with DMPK expanded-repeat transcripts.
abstract: Myotonic dystrophy (DM1) is the most common form of adult muscular dystrophy and is inherited as an autosomal dominant trait. The genetic basis of DM1 is the expansion of a CTG repeat in the 3' untranslated region of a protein kinase gene (DMPK). The molecular mechanism by which this expanded repeat produces the pathophysiology of DM1 remains unknown. Transcripts from the expanded allele accumulate as foci in the nucleus of DM1 cells and it has been suggested that these transcript foci sequester cellular proteins that are required for normal nuclear function. We have investigated the role of three RNA-binding proteins, CUG-BP, hnRNP C and MBNL, as possible sequestered factors. Using a combination of indirect immunofluorescence to detect endogenous proteins and overexpression of proteins with green fluorescent protein (GFP) tags we have shown that CUG-BP and hnRNP C do not co-localise with expanded repeat foci in DM1 cell lines. However, GFP-tagged MBNL does itself form foci in DM1 cell lines and co-localises with the foci of expanded repeat transcripts. GFP-tagged MBNL does not appear as foci in non-DM1 cell lines. This work provides further support for the involvement of MBNL in DM1.
authors: Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK.
authors: M
authors: M
authors: Fardaei
authors: K
authors: K
authors: Larkin
authors: J D
authors: JD
authors: Brook
authors: M G
authors: MG
authors: Hamshere
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000067879:CELF1 Protein; D005347:Fibroblasts; D019084:Fluorescent Antibody Technique, Indirect; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D017403:In Situ Hybridization; D008856:Microscopy, Fluorescence; D009223:Myotonic Dystrophy / Q000235:genetics*; D065636:Myotonin-Protein Kinase; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D021381:Protein Transport; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D019680:Trinucleotide Repeat Expansion / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; D000067879:CELF1 Protein; C102843:CELF1 protein, human; C585517:DMPK protein, human; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D065636:Myotonin-Protein Kinase; D017346:Protein Serine-Threonine Kinases