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Interaction between NBS1 and the mTOR/Rictor/SIN1 complex through specific domains.

Wang JQ, Chen JH, Chen YC, Chen MY, Hsieh CY, Teng SC, Wu KJ - PLoS ONE (2013)

Bottom Line: Knockdown of NBS1 decreased the levels of phosphorylated Akt and its downstream targets.Ionizing radiation (IR) increased the NBS1 levels and activated Akt activity.These results demonstrate that NBS1 interacts with the mTOR/Rictor/SIN1 complex through the a.a. 221-402 domain and contributes to the activation of Akt activity.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.

ABSTRACT
Nijmegen breakage syndrome (NBS) is a chromosomal-instability syndrome. The NBS gene product, NBS1 (p95 or nibrin), is a part of the Mre11-Rad50-NBS1 complex. SIN1 is a component of the mTOR/Rictor/SIN1 complex mediating the activation of Akt. Here we show that NBS1 interacted with mTOR, Rictor, and SIN1. The specific domains of mTOR, Rictor, or SIN1 interacted with the internal domain (a.a. 221-402) of NBS1. Sucrose density gradient showed that NBS1 was located in the same fractions as the mTOR/Rictor/SIN1 complex. Knockdown of NBS1 decreased the levels of phosphorylated Akt and its downstream targets. Ionizing radiation (IR) increased the NBS1 levels and activated Akt activity. These results demonstrate that NBS1 interacts with the mTOR/Rictor/SIN1 complex through the a.a. 221-402 domain and contributes to the activation of Akt activity.

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siRNA mediated knockdown of NBS1 decreased the levels of phosphorylated Akt (pAkt Ser-473) and its downstream targets and IR increased the levels of NBS1 and phosphorylated Akt.A. Western blot analysis showed that knockdown of NBS1 decreased the phosphorylated levels of Akt, GSK-3β, and Foxo1/3a). B. Ionizing radiation of two different cell lines (H1299, OCEM-1) increased the levels of NBS1 and phosphorylated Akt. C. Knockdown of NBS1 in H1299 cells abolished the increase in NBS1 and phosphorylated Akt levels under IR.
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pone-0065586-g006: siRNA mediated knockdown of NBS1 decreased the levels of phosphorylated Akt (pAkt Ser-473) and its downstream targets and IR increased the levels of NBS1 and phosphorylated Akt.A. Western blot analysis showed that knockdown of NBS1 decreased the phosphorylated levels of Akt, GSK-3β, and Foxo1/3a). B. Ionizing radiation of two different cell lines (H1299, OCEM-1) increased the levels of NBS1 and phosphorylated Akt. C. Knockdown of NBS1 in H1299 cells abolished the increase in NBS1 and phosphorylated Akt levels under IR.

Mentions: In order to test the correlation between NBS1 levels and its effect on the Akt activity, siRNA experiment to knockdown NBS1 was performed. The result showed that knockdown of NBS1 decreased the phosphorylated Akt levels (pAkt Ser-473) (Fig. 6), indicating that NBS1 contributed to the Akt activity. In addition, the phosphorylation levels of certain Akt downstream targets such as GSK-3β and Foxo1/3a were also decreased following NBS1 knockdown (Fig. 6A). These results supported the role of NBS1 in the activation of Akt activity.


Interaction between NBS1 and the mTOR/Rictor/SIN1 complex through specific domains.

Wang JQ, Chen JH, Chen YC, Chen MY, Hsieh CY, Teng SC, Wu KJ - PLoS ONE (2013)

siRNA mediated knockdown of NBS1 decreased the levels of phosphorylated Akt (pAkt Ser-473) and its downstream targets and IR increased the levels of NBS1 and phosphorylated Akt.A. Western blot analysis showed that knockdown of NBS1 decreased the phosphorylated levels of Akt, GSK-3β, and Foxo1/3a). B. Ionizing radiation of two different cell lines (H1299, OCEM-1) increased the levels of NBS1 and phosphorylated Akt. C. Knockdown of NBS1 in H1299 cells abolished the increase in NBS1 and phosphorylated Akt levels under IR.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3675082&req=5

pone-0065586-g006: siRNA mediated knockdown of NBS1 decreased the levels of phosphorylated Akt (pAkt Ser-473) and its downstream targets and IR increased the levels of NBS1 and phosphorylated Akt.A. Western blot analysis showed that knockdown of NBS1 decreased the phosphorylated levels of Akt, GSK-3β, and Foxo1/3a). B. Ionizing radiation of two different cell lines (H1299, OCEM-1) increased the levels of NBS1 and phosphorylated Akt. C. Knockdown of NBS1 in H1299 cells abolished the increase in NBS1 and phosphorylated Akt levels under IR.
Mentions: In order to test the correlation between NBS1 levels and its effect on the Akt activity, siRNA experiment to knockdown NBS1 was performed. The result showed that knockdown of NBS1 decreased the phosphorylated Akt levels (pAkt Ser-473) (Fig. 6), indicating that NBS1 contributed to the Akt activity. In addition, the phosphorylation levels of certain Akt downstream targets such as GSK-3β and Foxo1/3a were also decreased following NBS1 knockdown (Fig. 6A). These results supported the role of NBS1 in the activation of Akt activity.

Bottom Line: Knockdown of NBS1 decreased the levels of phosphorylated Akt and its downstream targets.Ionizing radiation (IR) increased the NBS1 levels and activated Akt activity.These results demonstrate that NBS1 interacts with the mTOR/Rictor/SIN1 complex through the a.a. 221-402 domain and contributes to the activation of Akt activity.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.

ABSTRACT
Nijmegen breakage syndrome (NBS) is a chromosomal-instability syndrome. The NBS gene product, NBS1 (p95 or nibrin), is a part of the Mre11-Rad50-NBS1 complex. SIN1 is a component of the mTOR/Rictor/SIN1 complex mediating the activation of Akt. Here we show that NBS1 interacted with mTOR, Rictor, and SIN1. The specific domains of mTOR, Rictor, or SIN1 interacted with the internal domain (a.a. 221-402) of NBS1. Sucrose density gradient showed that NBS1 was located in the same fractions as the mTOR/Rictor/SIN1 complex. Knockdown of NBS1 decreased the levels of phosphorylated Akt and its downstream targets. Ionizing radiation (IR) increased the NBS1 levels and activated Akt activity. These results demonstrate that NBS1 interacts with the mTOR/Rictor/SIN1 complex through the a.a. 221-402 domain and contributes to the activation of Akt activity.

Show MeSH
Related in: MedlinePlus