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Originally published In Press as doi:10.1074/jbc.M206201200 on September 9, 2002

J. Biol. Chem., Vol. 277, Issue 47, 45480-45492, November 22, 2002
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AAT-1, a Novel Testis-specific AMY-1-binding Protein, Forms a Quaternary Complex with AMY-1, A-kinase Anchor Protein 84, and a Regulatory Subunit of cAMP-dependent Protein Kinase and Is Phosphorylated by Its Kinase*

Hiroshi YukitakeDagger , Makoto FurusawaDagger §, Takahiro TairaDagger §, Sanae M. M. Iguchi-Ariga§, and Hiroyoshi ArigaDagger §||

From the Dagger  Graduate School of Pharmaceutical Sciences,  College of Medical Technology, Hokkaido University, Kita-ku, Sapporo 060-0812, Japan, and § CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan

AMY-1 has been identified by us as a c-Myc-binding protein and was found to stimulate c-Myc transcription activity. AMY-1 was also found to be associated with protein kinase A anchor protein 84/149 (S-AKAP84/AKAP149) in the mitochondria in somatic cells and sperm, suggesting that it plays a role in spermatogenesis. To determine the molecular function of AMY-1, a two-hybrid screening of cDNAs encoding AMY-1-binding proteins was carried out with AMY-1 as a bait using a human testis cDNA library, and a clone encoding a novel protein, AAT-1, was obtained. Three isoforms of AAT-1, AAT-1alpha , -beta , and -gamma , were found to be derived from an alternative splicing of the transcripts of the aat-1 gene, which was mapped at human chromosome 3q13-3q21. AAT-1 was found to be specifically expressed in the testis during the course of spermatogenesis and also to be present in the spermatid and mature sperm, as was AMY-1. AAT-1alpha was found to bind to and be colocalized in mitochondria with AMY-1 in human HeLa and mouse GC-1 cells. Furthermore, AAT-1alpha was found to bind to the N-terminal half of S-AKAP84/AKAP149 in a quaternary complex with AMY-1 and a regulatory subunit (RII) of cAMP-dependent kinase (PKA), in which AAT-1alpha was associated with RII via S-AKAP84/AKAP149, in rat testis and HeLa cells. It was then found that AAT-1alpha weakly stimulated a phosphorylation activity of PKA and also that AAT-1 itself was phosphorylated by PKA in vivo and in vitro. These results suggest that both AAT-1 and AMY-1 play roles in spermatogenesis.


* This work was supported by grants-in-aid from the Ministry of Education, Science, Culture and Sport 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) AB063296-AB063298.

|| To whom correspondence should be addressed: Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita 12, Nishi 6, Kita-ku, Sapporo 060-0812, Japan. Tel.: 81-11-706-3745; Fax: 81-11-706-4988; E-mail: hiro@pharm.hokudai.ac.jp.


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