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mTOR-rictor is the Ser473 kinase for AKT1 in mouse one-cell stage embryos

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Abstract

Mammalian target of rapamycin (mTOR) controls cell growth and proliferation via the raptor-mTOR (TORC1) and rictor-mTOR (TORC2) protein complexes. The mTORC2 containing mTOR and rictor is thought to be rapamycin insensitive and it is recently shown that both rictor and mTORC2 are essential for the development of both embryonic and extra embryonic tissues. To explore rictor function in the early development of mouse embryos, we disrupted the expression of rictor, a specific component of mTORC2, in mouse fertilized eggs by using rictor shRNA. Our results showed that one-cell stage eggs that were lack of rictor could not enter into the two-cell stage normally. Recent biochemical studies suggests that TORC2 is the elusive PDK2 (3′-phosphoinositide-dependent kinase 2) for AKT/PKB Ser473 phosphorylation, which is deemed necessary for AKT function, so we microinjected AKT-S473A into mouse fertilized eggs to investigate whether AKT-S473A is downstream effector of mTOR.rictor to regulate the mitotic division. Our findings revealed that the rictor induced phosphorylation of AKT in Ser473 is required for TORC2 function in early development of mouse embryos.

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Acknowledgments

We thank Dr. Estella Jacinto (University of Basel, Switzerland) for presenting the constructs of raptor shRNA and rictor shRNA as a gift. We also thank PhD. Chen Feng (Dept. of biochemistry and molecular biology, China Medical University, China) for offering the wild-type AKT1 plasmid (pBS-AKT1-WT).

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Authors and Affiliations

  1. Department of Urology, No. 1 Hospital of China Medical University, Shenyang, Liaoning, People’s Republic of China

    Zhe Zhang

  2. Department of Hematology, Shengjing Hospital of China Medical University, Shenyang, Liaoning, People’s Republic of China

    Guojun Zhang

  3. Department of Pathophysiology, College of Basic Medicine of China Medical University, Shenyang, Liaoning, People’s Republic of China

    Xiaoyan Xu

  4. Department of Biochemical and Molecular Biology, China Medical University, Beier Road, Heping District, Shenyang, Liaoning Province, 110001, People’s Republic of China

    Zhe Zhang, Wenhui Su & Bingzhi Yu

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  1. Zhe Zhang
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  2. Guojun Zhang
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  3. Xiaoyan Xu
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  4. Wenhui Su
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  5. Bingzhi Yu
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Corresponding author

Correspondence to Bingzhi Yu.

Additional information

The study was supported by National Natural Science Foundation of China (81070489).

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Zhang, Z., Zhang, G., Xu, X. et al. mTOR-rictor is the Ser473 kinase for AKT1 in mouse one-cell stage embryos. Mol Cell Biochem 361, 249–257 (2012). https://doi.org/10.1007/s11010-011-1110-0

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