In cultured oligodendrocytes the A/B-type hnRNP CBF-A accompanies MBP mRNA bound to mRNA trafficking sequences

Chandrasekhar S. Raju, Christian Göritz, Ylva Nord, Ola Hermanson, Carmen López-Iglesias, Neus Visa, Goncalo Castelo-Branco, Piergiorgio Percipalle

Research output: Contribution to journalArticle

Abstract

Heterogeneous ribonucleoproteins (hnRNPs) have key roles in RNA biogenesis, including pre-mRNP assembly, transport and cytoplasmic localization. Here we show by biochemical fractionation of nuclear extracts and protein-protein interaction assays that the A/B-type hnRNP CBF-A is in a multiprotein complex with hnRNP A2 and A3 and hnRNP U. Using RNA affinity chromatography and gel retardation assays, CBF-A was found to bind directly to RNA trafficking sequences in the 3′-UTR of the myelin basic protein (MBP) mRNA. In primary oligodendrocytes, astrocytes, neurons, and mouse forebrain sections, CBF-A revealed a characteristic granular cytoplasmic distribution. In mouse forebrain CBF-A-positive granules were preferentially found in regions with loosely bundled myelin fibers. In cultured oligodendrocytes, CBF-A was found to be specifically associated with endogenous MBP mRNA and CBF-A gene silencing resulted in the retention of MBP granules in the cell body. Finally, immunoelectron microscopy in differentiating oligodendrocytes showed that CBF-A is located in cytoplasmic granules that are often associated with the cytoskeleton. The results suggest that CBF-A is a novel transacting factor required for cytoplasmic mRNA transport and localization.

Original languageEnglish (US)
Pages (from-to)3008-3019
Number of pages12
JournalMolecular Biology of the Cell
Volume19
Issue number7
DOIs
StatePublished - Jul 1 2008

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Heterogeneous-Nuclear Ribonucleoproteins
Myelin Basic Protein
Oligodendroglia
Prosencephalon
Messenger RNA
Heterogeneous-Nuclear Ribonucleoprotein U
RNA
Cytoplasmic Granules
Multiprotein Complexes
Ribonucleoproteins
Immunoelectron Microscopy
3' Untranslated Regions
Gene Silencing
Electrophoretic Mobility Shift Assay
Myelin Sheath
Nuclear Proteins
Cytoskeleton
Affinity Chromatography
Astrocytes
Neurons

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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In cultured oligodendrocytes the A/B-type hnRNP CBF-A accompanies MBP mRNA bound to mRNA trafficking sequences. / Raju, Chandrasekhar S.; Göritz, Christian; Nord, Ylva; Hermanson, Ola; López-Iglesias, Carmen; Visa, Neus; Castelo-Branco, Goncalo; Percipalle, Piergiorgio.

In: Molecular Biology of the Cell, Vol. 19, No. 7, 01.07.2008, p. 3008-3019.

Research output: Contribution to journalArticle

Raju, CS, Göritz, C, Nord, Y, Hermanson, O, López-Iglesias, C, Visa, N, Castelo-Branco, G & Percipalle, P 2008, 'In cultured oligodendrocytes the A/B-type hnRNP CBF-A accompanies MBP mRNA bound to mRNA trafficking sequences', Molecular Biology of the Cell, vol. 19, no. 7, pp. 3008-3019. https://doi.org/10.1091/mbc.E07-10-1083
Raju, Chandrasekhar S. ; Göritz, Christian ; Nord, Ylva ; Hermanson, Ola ; López-Iglesias, Carmen ; Visa, Neus ; Castelo-Branco, Goncalo ; Percipalle, Piergiorgio. / In cultured oligodendrocytes the A/B-type hnRNP CBF-A accompanies MBP mRNA bound to mRNA trafficking sequences. In: Molecular Biology of the Cell. 2008 ; Vol. 19, No. 7. pp. 3008-3019.
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