Identification and analysis of function of a novel splicing variant of mouse receptor activator of NF-κB

Satomi Mukai, Riko Kitazawa, Junko Ishii, Takeshi Kondo, Akihiro Hakozaki, Keisuke Horiuchi, Ryuma Haraguch, Kiyoshi Mori, Sohei Kitazawa

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Receptor activator of NF-κB (RANK) is a member of the tumor necrosis factor receptor (TNFR) family expressed in osteoclast precursors, and RANK-RANK ligand (RANKL) signaling is a key system for differentiation, activation and survival of osteoclasts. Here, we report the identification of a novel alternative splicing variant of mouse RANK gene (vRANK) that contains a new intervening exon between exon 1 and exon 2 of mouse full-length RANK (fRANK) mRNA. Since this novel exon contains the stop codon, vRANK encodes truncated amino acids that have a portion of the signal peptide of fRANK and an additional 19 amino acids that show no homology to previously reported domains. By transient transfection studies with vRANK-GFP and -Flag expressing constructs, vRANK was found localized mostly in the cytoplasm and partly in the cell membrane, but was not secreted into the culture supernatant. Under the stimulation of various factors, the expression of vRANK mRNA was almost parallel to that of fRANK in RAW264.7 cells not treated with M-CSF. Overexpression of vRANK, on the other hand, decreased TRACP (a marker of osteoclasts) mRNA expression as well as the number of TRACP-positive multinucleated giant cells. While the mRNA expression levels of NFATc1 (a master transcriptional factor of the osteoclast differentiation program) were not affected, apoptotic cells increased significantly in vRAN K-transfected cells treated with sRANKL. Taken together, these results suggest that vRANK is a novel osteoclast suppressor that reduces the number of RANKL-induced mature osteoclasts mainly by negating the anti-apoptotic effect of RANKL.

Original languageEnglish
Pages (from-to)29-38
Number of pages10
JournalMolecular and Cellular Biochemistry
Volume350
Issue number1-2
DOIs
Publication statusPublished - 2011 Apr 1

Keywords

  • Alternative splicing
  • Osteoclast
  • RANK
  • RAW264.7

ASJC Scopus subject areas

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

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  • Cite this

    Mukai, S., Kitazawa, R., Ishii, J., Kondo, T., Hakozaki, A., Horiuchi, K., Haraguch, R., Mori, K., & Kitazawa, S. (2011). Identification and analysis of function of a novel splicing variant of mouse receptor activator of NF-κB. Molecular and Cellular Biochemistry, 350(1-2), 29-38. https://doi.org/10.1007/s11010-010-0679-z