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Journal of Bone and Mineral Metabolism

, Volume 36, Issue 4, pp 410–419 | Cite as

THRAP3 interacts with and inhibits the transcriptional activity of SOX9 during chondrogenesis

  • Takashi Sono
  • Haruhiko AkiyamaEmail author
  • Shigenori Miura
  • Jian Min Deng
  • Chisa Shukunami
  • Yuji Hiraki
  • Yu Tsushima
  • Yoshiaki Azuma
  • Richard R. Behringer
  • Shuichi Matsuda
Original Article

Abstract

Sex-determining region Y (Sry)-box (Sox)9 is required for chondrogenesis as a transcriptional activator of genes related to chondrocyte proliferation, differentiation, and cartilage-specific extracellular matrix. Although there have been studies investigating the Sox9-dependent transcriptional complexes, not all their components have been identified. In the present study, we demonstrated that thyroid hormone receptor-associated protein (THRAP)3 is a component of a SOX9 transcriptional complex by liquid chromatography mass spectrometric analysis of FLAG-tagged Sox9-binding proteins purified from FLAG-HA-tagged Sox9 knock-in mice. Thrap3 knockdown in ATDC5 chondrogenic cells increased the expression of Collagen type II alpha 1 chain (Col2a1) without affecting Sox9 expression. THRAP3 and SOX9 overexpression reduced Col2a1 levels to a greater degree than overexpression of SOX9 alone. The negative regulation of SOX9 transcriptional activity by THRAP3 was mediated by interaction between the proline-, glutamine-, and serine-rich domain of SOX9 and the innominate domain of THRAP3. These results indicate that THRAP3 negatively regulates SOX9 transcriptional activity as a cofactor of a SOX9 transcriptional complex during chondrogenesis.

Keywords

THRAP3 SOX9 Chondrogenesis Knock-in mouse LC/MS/MS 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they are no conflict of interest.

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Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan KK 2017

Authors and Affiliations

  • Takashi Sono
    • 1
  • Haruhiko Akiyama
    • 2
    Email author
  • Shigenori Miura
    • 3
  • Jian Min Deng
    • 4
  • Chisa Shukunami
    • 3
    • 5
  • Yuji Hiraki
    • 3
  • Yu Tsushima
    • 6
  • Yoshiaki Azuma
    • 7
  • Richard R. Behringer
    • 4
  • Shuichi Matsuda
    • 1
  1. 1.Department of Orthopaedic Surgery, Graduate School of MedicineKyoto UniversityKyotoJapan
  2. 2.Department of Orthopaedic SurgeryGifu UniversityGifuJapan
  3. 3.Laboratory of Cellular Differentiation, Institute for Frontier Life and Medical SciencesKyoto UniversityKyotoJapan
  4. 4.Department of GeneticsUniversity of Texas M.D. Anderson Cancer CenterHoustonUSA
  5. 5.Department of Molecular Biology and Biochemistry, Division of Basic Life Sciences, Institute of Biomedical and Health SciencesHiroshima UniversityHiroshimaJapan
  6. 6.Pharmacology Research DepartmentTeijin Pharma LimitedHinoJapan
  7. 7.Medical Science DepartmentTeijin Pharma LimitedTokyoJapan

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