Nucleophosmin/B23: A multifunctional regulator that determines the fate of CCN2 mRNA

Satoshi Kubota, Yoshiki Mukudai, Harumi Kawaki, Seiji Kondo, Takanori Eguchi, Kumi Sumiyoshi, Toshihiro Ohgawara, Tsuyoshi Shimo, Masaharu Takigawa

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

CCN2/CTGF is a multifunctional molecule that has been shown to play a central role in chondrocyte differentiation. During this process, the expression of ccn2 is tightly regulated to confer a maximal level at prehypertrophic - hypertrophic stages, in which the 3'-untranslated region (UTR) of the mRNA is critically involved in mediating its post-transcriptional regulation. In our previous studies, we found that a 40-kDa protein binding specifically to an RNA cis-element, 3'-100/50, in the 3'-UTR of the chicken ccn2 mRNA regulated the intracellular stability of the mRNA. The interaction of this 40-kDa protein with 3-100/50 was enhanced in proliferating chondrocytes, in which ccn2 mRNA is rapidly degraded; whereas a prolonged half life of ccn2 mRNA is observed in hypertrophic chondrocytes, where the interaction of the 40 kDa-protein and 3'-100/50 is diminished. Collectively, the data suggested that this 40-kDa protein acts as a ccn2-specific mRNA destabilizer during chondrocyte differentiation. In this present study we finally identified this 40-kDa protein as nucleophosmin (NPM)/B23. NPM is a nuclear-cytoplasmic shuttling protein that is characterized by its multiple functionality. This protein is known to be a histone chaperone, a regulator of ribosomal RNA transcription, as well as an RNA-binding post-transcriptional regulator of gene expression. In our hands, direct binding of NPM to 3'-100/50 was confirmed not only by RNA EMSA and UV crosslinking assays, but also by RNA immunoprecipitation analysis. By using recombinant chicken NPM, we could successfully reconstitute the post-transcriptional regulation of ccn2 by NPM in vitro and found that this regulation was more robust in chondrocytes than in fibroblasts. Furthermore, siRNA-mediated gene silencing of NPM in vivo clearly showed enhanced ccn2 gene expression and a prolonged half life of the ccn2 mRNA, confirming the functional property of NPM as a specific destabilizer of the ccn2 mRNA in living cells. The 5'-100/50 element, a target of NPM, is evolutionally conserved among vertebrate species. Therefore, we consider NPM to be a critical post-transcriptional regulator of ccn2 acting via 3'-UTR during endochondral ossification and possibly, in other physiological and pathological states as well.

Original languageEnglish
Title of host publicationCCN Proteins in Health and Disease
Subtitle of host publicationAn Overview of the Fifth International Workshop on the CCN Family of Genes
PublisherSpringer Netherlands
Pages41-55
Number of pages15
ISBN (Print)9789048137787
DOIs
Publication statusPublished - 2010

Keywords

  • B23
  • CCN2
  • Chondrocytes
  • Nucleophosmin
  • Post-transcriptional regulation

ASJC Scopus subject areas

  • Medicine(all)

Fingerprint Dive into the research topics of 'Nucleophosmin/B23: A multifunctional regulator that determines the fate of CCN2 mRNA'. Together they form a unique fingerprint.

  • Cite this

    Kubota, S., Mukudai, Y., Kawaki, H., Kondo, S., Eguchi, T., Sumiyoshi, K., Ohgawara, T., Shimo, T., & Takigawa, M. (2010). Nucleophosmin/B23: A multifunctional regulator that determines the fate of CCN2 mRNA. In CCN Proteins in Health and Disease: An Overview of the Fifth International Workshop on the CCN Family of Genes (pp. 41-55). Springer Netherlands. https://doi.org/10.1007/978-90-481-3779-4_4