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Post‐transcriptional regulation of fukutin in an astrocytoma cell line
Author(s) -
Yamamoto Tomoko,
Kato Yoichiro,
Hiroi Atsuko,
Shibata Noriyuki,
Osawa Makiko,
Kobayashi Makio
Publication year - 2012
Publication title -
international journal of experimental pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.671
H-Index - 72
eISSN - 1365-2613
pISSN - 0959-9673
DOI - 10.1111/j.1365-2613.2011.00799.x
Subject(s) - microbiology and biotechnology , biology , gene knockdown , messenger rna , in situ hybridization , cell culture , gene , genetics
Summary Fukutin is the gene responsible for Fukuyama‐type congenital muscular dystrophy (FCMD), an autosomal recessive disease associated with central nervous system (CNS) and eye anomalies. Fukutin is involved in basement membrane formation via the glycosylation of α‐dystroglycan (α‐DG), and hypoglycosylation of α‐DG provokes the muscular, CNS and eye lesions of FCMD. Astrocytes play an important role in the pathogenesis of the CNS lesions, but the post‐transcriptional regulation of fukutin mRNA has not been elucidated. In this study, we investigated the characteristics of fukutin mRNA using an astrocytoma cell line that expresses fukutin and glycosylated α‐DG. The glycosylation of α‐DG was considered to be increased by over‐expression of fukutin and decreased by knockdown of fukutin. Knockdown of Musashi‐1, one of the RNA‐binding proteins involved in the regulation of neuronal differentiation, induced a decrease in fukutin mRNA. Immunoprecipitation and ELISA‐based RNA‐binding assay demonstrated possible binding between fukutin mRNA and Musashi‐1 protein. A relationship between fukutin mRNA and vimentin protein was also proposed. In situ hybridization for fukutin mRNA showed a positive cytoplasmic reaction including cytoplasmic processes. From these results, fukutin mRNA is suggested to be a localized mRNA up‐regulated by Musashi‐1 and to be a component of a mRNA‐protein complex which includes Musashi‐1 and (presumably) vimentin proteins.