A novel protein kinase B (PKB)/AKT-binding protein enhances PKB kinase activity and regulates DNA synthesis

Motonobu Anai, Nobuhiro Shojima, Hideki Katagiri, Takehide Ogihara, Hideyuki Sakoda, Yukiko Onishi, Hiraku Ono, Midori Fujishiro, Yasushi Fukushima, Nanao Horike, Amelia Viana, Masatoshi Kikuchi, Noriko Noguchi, Shinichiro Takahashi, Kuniaki Takata, Yoshitomo Oka, Yasunobu Uchijima, Hiroki Kurihara, Tomoichiro Asano

Research output: Contribution to journalArticlepeer-review

103 Citations (Scopus)

Abstract

Protein kinase B (PKB)/Akt reportedly plays a role in the survival and/or proliferation of cells. We identified a novel protein, which binds to PKB, using a yeast two-hybrid screening system. This association was demonstrated not only in vivo by overexpressing both proteins or by coimmunoprecipitation of the endogenous proteins, but also in vitro using glutathione S-transferase fusion proteins. Importantly, this protein specifically associates with the C terminus of PKB but not with other AGC kinases and enhances PKB phosphorylation and kinase activation without growth factor stimulation. Thus, we termed this Akt-specific binding protein APE (Akt-phosphorylation enhancer). Since APE-induced phosphorylation of PKB did not occur in cells treated with wortmannin or LY294002, APE itself is not a kinase but seems to enhance or prolong the phosphoinositide 3-kinase-dependent phosphorylation of PKB. In cells in which APE was suppressed by small interfering RNA, DNA synthesis was significantly reduced with suppression of PKB phosphorylation, suggesting a synergistic role of APE in PKB-induced proliferation. On the other hand, in cells overexpressing both PKB and APE, despite markedly increased basal phosphorylation of PKB, both DNA rereplication and subsequent Chk2 phosphorylation and apoptosis were seen, suggesting the involvement of APE in the regulation of cell cycling replication licensing. Taking these observations together, APE appears to be a novel regulator of PKB phosphorylation. Furthermore, the interaction between APE and PKB, possibly dependent on the expression levels of both proteins, may be a novel molecular mechanism leading to proliferation and/or apoptosis.

Original languageEnglish
Pages (from-to)18525-18535
Number of pages11
JournalJournal of Biological Chemistry
Volume280
Issue number18
DOIs
Publication statusPublished - 2005 May 6
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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