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Originally published In Press as doi:10.1074/jbc.M104719200 on November 5, 2001

J. Biol. Chem., Vol. 277, Issue 2, 1443-1450, January 11, 2002
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Functional Analysis of the Chondroitin 6-Sulfotransferase Gene in Relation to Lymphocyte Subpopulations, Brain Development, and Oversulfated Chondroitin Sulfates*

Kenji UchimuraDagger §, Kenji KadomatsuDagger , Hitoshi Nishimura, Hisako MuramatsuDagger , Eishin NakamuraDagger ||, Nobuyuki KurosawaDagger , Osami Habuchi**, Fathy M. El-FasakhanyDagger , Yasunobu Yoshikai, and Takashi MuramatsuDagger Dagger Dagger

From the Dagger  Department of Biochemistry, the  Laboratory of Host Defense & Germfree Life, Research Institute for Disease Mechanism and Control, and the || Department of Internal Medicine, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550 and the ** Department of Life Science, Aichi University of Education, Kariya, Aichi 448-8542, Japan

Chondroitin 6-sulfotransferase (C6ST) catalyzes the transfer of sulfate to position 6 of the N-acetylgalactosamine residue of chondroitin. To obtain direct evidence regarding the function of C6ST and its product, chondroitin 6-sulfate, in vivo, we isolated the mouse C6ST gene (C6st) and generated mice deficient in this gene (C6st-/-) by embryonic stem cell technology. C6st-/- mice were born at approximately the expected frequency and were viable through adulthood. In the spleen of C6st-/- mice, the level of chondroitin 6-sulfate became almost undetectable. Analyses of these knockout mice provided insights into the biosynthesis of oversulfated chondroitin sulfates in mice; chondroitin sulfate D in the brain of null mice and the cartilage and telencephalon of null embryos disappeared, whereas the chondroitin sulfate E level in the spleen and brain of the null mice was unchanged. Despite the disappearance of chondroitin sulfate D structure, brain development was normal in the C6st-/- mice. Further analysis revealed that the number of CD62L+CD44low T lymphocytes corresponding to naive T lymphocytes in the spleen of 5-6-week-old C6st-/- mice was significantly decreased, whereas those in other secondary lymphoid organs were unchanged. This finding suggested that chondroitin 6-sulfate plays a role in the maintenance of naive T lymphocytes in the spleen of young mice.

* This work was supported in part by Grants-in-aid for Scientific Research 12003898 and 12480187 from the Japan Society for the Promotion of Science and Grants-in-aid 09680616 and 10178102 for Scientific Research from the Ministry of Education, Science, Culture and Sports 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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AB062107-AB062109.

§ Research fellow of the Japan Society for the Promotion of Science.

Dagger Dagger To whom correspondence should be addressed. Tel.: 81-52-744-2059; Fax: 81-52-744-2065; E-mail: tmurama@med.nagoya-u.ac.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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