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J. Biol. Chem., Vol. 277, Issue 47, 45480-45492, November 22, 2002
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From the 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-1 The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AB063296-AB063298.
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*
,
§,
§,
§
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
,
-
, and -
, 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-1
was found to bind to and be colocalized in mitochondria with AMY-1 in
human HeLa and mouse GC-1 cells. Furthermore, AAT-1
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-1
was associated with RII via
S-AKAP84/AKAP149, in rat testis and HeLa cells. It was then found that
AAT-1
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.
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.
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