Nucleolar localization of DGCR8 and identification of eleven DGCR8-associated proteins

Aiko Shiohama, Takashi Sasaki, Setsuko Noda, Shinsei Minoshima, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

We identified 11 proteins that are associated with DGCR8 by immunoprecipitation assay and mass spectrometry. These proteins included Nucleolin, ILF3 and others, most of which appeared to be involved in the RNA processing or RNA transportation. We detected at least four kinds of protein complex, such as DROSHA/DGCR8, DGCR8/Nucleolin, DGCR8/ILF3 and ILF3/XPO5, by co-immunoprecipitation. The complex formation of DGCR8 with Nucleolin was dependent on RNA. Subcellular localization analysis by the immunofluorescent microscopy and immunoelectron microscopy indicated that DGCR8 locates at the nucleolus and small foci adjacent to splicing speckles in the nucleoplasm. Furthermore, the localization of DGCR8 at the nucleolus was changed by the inhibition of RNA transcription. Thus, our studies provided additional new evidence for the involvement of various protein complexes in the molecular mechanisms of apparently complex innate RNA interference machinery.

Original languageEnglish
Pages (from-to)4196-4207
Number of pages12
JournalExperimental Cell Research
Volume313
Issue number20
DOIs
Publication statusPublished - 2007 Dec 10

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RNA
Immunoprecipitation
Proteins
Immunoelectron Microscopy
RNA Interference
Microscopy
Mass Spectrometry
nucleolin

Keywords

  • Nuclear body
  • Nucleolus
  • Protein-protein interaction
  • RNA

ASJC Scopus subject areas

  • Cell Biology

Cite this

Nucleolar localization of DGCR8 and identification of eleven DGCR8-associated proteins. / Shiohama, Aiko; Sasaki, Takashi; Noda, Setsuko; Minoshima, Shinsei; Shimizu, Nobuyoshi.

In: Experimental Cell Research, Vol. 313, No. 20, 10.12.2007, p. 4196-4207.

Research output: Contribution to journalArticle

Shiohama, Aiko ; Sasaki, Takashi ; Noda, Setsuko ; Minoshima, Shinsei ; Shimizu, Nobuyoshi. / Nucleolar localization of DGCR8 and identification of eleven DGCR8-associated proteins. In: Experimental Cell Research. 2007 ; Vol. 313, No. 20. pp. 4196-4207.
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