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J. Biol. Chem., Vol. 279, Issue 11, 10765-10775, March 12, 2004

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Mutual Regulation of Protein-tyrosine Phosphatase 20 and Protein-tyrosine Kinase Tec Activities by Tyrosine Phosphorylation and Dephosphorylation^*

Naohito Aoki, Shuichi Ueno¶, Hiroyuki Mano¶, Sho Yamasaki||, Masayuki Shiota**, Hitoshi Miyazaki**, Yumiko Yamaguchi-Aoki, Tsukasa Matsuda, and Axel Ullrich

From the Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan, ^¶Divisions of Functional Genomics, Cardiology and Hematology, Jichi Medical School, Kawachi-gun, Tochigi 329-0498, Japan, ^||Molecular Genetics, Chiba University Graduate School of Medicine, Chiba 260-8670, Japan, ^**Gene Research Center, University of Tsukuba, Ibaraki 305-8572, Japan, and Max Planck Institute for Biochemistry, Department of Molecular Biology, D-82152 Martinsried, Germany

PTP20, also known as HSCF/protein-tyrosine phosphatase K1/fetal liver phosphatase 1/brain-derived phosphatase 1, is a cytosolic protein-tyrosine phosphatase with currently unknown biological relevance. We have identified that the nonreceptor protein-tyrosine kinase Tec-phosphorylated PTP20 on tyrosines and co-immunoprecipitated with the phosphatase in a phosphotyrosine-dependent manner. The interaction between the two proteins involved the Tec SH2 domain and the C-terminal tyrosine residues Tyr-281, Tyr-303, Tyr-354, and Tyr-381 of PTP20, which were also necessary for tyrosine phosphorylation/dephosphorylation. Association between endogenous PTP20 and Tec was also tyrosine phosphorylation-dependent in the immature B cell line Ramos. Finally, the Tyr-281 residue of PTP20 was shown to be critical for deactivating Tec in Ramos cells upon B cell receptor ligation as well as dephosphorylation and deactivation of Tec and PTP20 itself in transfected COS7 cells. Taken together, PTP20 appears to play a negative role in Tec-mediated signaling, and Tec-PTP20 interaction might represent a negative feedback mechanism.

Received for publication, September 22, 2003 , and in revised form, December 5, 2003.

^* This work was supported in part by grants-in-aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan (to N. A. and T. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom all correspondence should be addressed: Dept. of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan. Fax: 81-52-789-4128; E-mail: naoki{at}agr.nagoya-u.ac.jp.

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