Identification of Type I and Type II Serine/Threonine Kinase Receptors for Growth/Differentiation Factor-5

doi:10.1074/jbc.271.35.21345

Identification of Type I and Type II Serine/Threonine Kinase Receptors for Growth/Differentiation Factor-5*

From the ^‡ Department of Biochemistry, the Cancer Institute, Tokyo, Japanese Foundation for Cancer Research, 1-37-1 Kami-Ikebukuro, Toshima-ku, Tokyo 170, Japan, the
^§ Second Department of Oral and Maxillofacial Surgery, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113, Japan,
^∥ Drug Discovery Research Laboratories, Pharma Research & Development Division, Hoechst Japan Limited, 1-3-2 Minamidai, Kawagoe 350-11, Japan, the
^″ Department of Oral Pathology, School of Dentistry, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142, Japan, and the
^‴ Department of Ophthalmology, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113, Japan

^¶ To whom correspondence should be addressed. Tel./Fax: 81-3-3918-0342.

Abstract

Growth/differentiation factor-5 (GDF-5) is a member of the bone morphogenetic protein (BMP) family, which plays an important role in bone development in vivo. Mutations in the GDF-5 gene result in brachypodism in mice and Hunter-Thompson type chondrodysplasia in human. BMPs transduce their effects through binding to two different types of serine/threonine kinase receptors, type I and type II. However, binding abilities appear to be different among the members of the BMP family. BMP-4 binds to two different type I receptors, BMP receptors type IA (BMPR-IA) and type IB (BMPR-IB), and a type II receptor, BMP receptor type II (BMPR-II). In addition to these receptors, osteogenic protein-1 (OP-1, also known as BMP-7) binds to activin type I receptor (ActR-I) as well as activin type II receptors (ActR-II and ActR-IIB). Here we investigate the binding and signaling properties of GDF-5 through type I and type II receptors. GDF-5 induced alkaline phosphatase activity in a rat osteoprogenitor-like cell line, ROB-C26. ¹²⁵I-GDF-5 bound to BMPR-IB and BMPR-II but not to BMPR-IA in ROB-C26 cells and other nontransfected cell lines. Analysis using COS-1 cells transfected with the receptor cDNAs revealed that GDF-5 bound to BMPR-IB but not to the other type I receptors when expressed alone. When COS-1 cells were transfected with type II receptor cDNAs, GDF-5 bound to ActR-II, ActR-IIB, and BMPR-II but not to transforming growth factor-β type II receptor. In the presence of type II receptors, GDF-5 bound to different sets of type I receptors, but the binding was most efficient to BMPR-IB compared with the other type I receptors. Moreover, a transcriptional activation signal was efficiently transduced by BMPR-IB in the presence of BMPR-II or ActR-II after stimulation by GDF-5. These results suggest that BMPR-IB mediates certain signals for GDF-5 after forming the heteromeric complex with BMPR-II or ActR-II.

Footnotes

↵^∥ Supported by the Japan Research Foundation for Clinical Pharmacology.
↵* This work was supported in part by grants-in-aid for scientific research from the Ministry of Education, Science and Culture of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

GDF

growth/differentiation factor

CDMP

cartilage-derived morphogenetic protein

BMP

bone morphogenetic protein

TGF-β

transforming growth factor-β

DPP

decapentaplegic gene product

OP

osteogenic protein

ALK

activin receptor-like kinase

TβR

TGF-β

receptor

ActR

activin receptor

BMPR

BMP receptor

FBS

fetal bovine serum

ALP

alkaline phosphatase

NTA

nitrilotriacetic acid.
↵² H. Nishitoh, M. Saitoh, I. Asahina, S. Enomoto, T. K. Sampath, M. Takagi, and H. Ichijo, submitted for publication.
↵³ M. Takahashi, H. Tanaka, S. Kawai, H. Pan, K. Osawa, H. Miki, Y. Muraki, Y. Konno, T. Itoh, H. Takamatsu, K. Enomoto, M. Katsuura, and M. Kimura, manuscript in preparation.
↵⁴ H. Nishitoh, H. Ichijo, M. Kimura, T. Matsumoto, F. Makishima, M. Kato, and K. Miyazono, unpublished observations.
- Received April 22, 1996.