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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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Mapping of the domain in NBS1 interacting with mTOR or Rictor.A & B. Co-immunoprecipitation assays showed the interaction of the internal domain (a.a. 221-402) of NBS1 with mTOR. C & D. Co-immunoprecipitation assays showed the interaction of the internal domain of NBS1 with Rictor.
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pone-0065586-g003: Mapping of the domain in NBS1 interacting with mTOR or Rictor.A & B. Co-immunoprecipitation assays showed the interaction of the internal domain (a.a. 221-402) of NBS1 with mTOR. C & D. Co-immunoprecipitation assays showed the interaction of the internal domain of NBS1 with Rictor.

Mentions: In order to map the domain in NBS1 that interacts with mTOR or Rictor, different truncation mutants of NBS1 (C-terminal truncation mutant NBS1-653 and N-terminal truncation mutant NBSp70) were co-expressed with mTOR or Rictor followed by co-immunoprecipitation [12]. The result showed that both the mutants interacted with mTOR and Rictor (Fig. 3A, C). Further fine mapping of the domain using two different NBS1 truncation mutants (NBS221-402 and NBS402-653) together with mTOR or Rictor showed that only NBS221-402 interacted with mTOR and Rictor (Fig. 3B, D). All the results indicated that the internal domain (a.a. 221-402) of NBS1 interacted with both mTOR and Rictor.


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)

Mapping of the domain in NBS1 interacting with mTOR or Rictor.A & B. Co-immunoprecipitation assays showed the interaction of the internal domain (a.a. 221-402) of NBS1 with mTOR. C & D. Co-immunoprecipitation assays showed the interaction of the internal domain of NBS1 with Rictor.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0065586-g003: Mapping of the domain in NBS1 interacting with mTOR or Rictor.A & B. Co-immunoprecipitation assays showed the interaction of the internal domain (a.a. 221-402) of NBS1 with mTOR. C & D. Co-immunoprecipitation assays showed the interaction of the internal domain of NBS1 with Rictor.
Mentions: In order to map the domain in NBS1 that interacts with mTOR or Rictor, different truncation mutants of NBS1 (C-terminal truncation mutant NBS1-653 and N-terminal truncation mutant NBSp70) were co-expressed with mTOR or Rictor followed by co-immunoprecipitation [12]. The result showed that both the mutants interacted with mTOR and Rictor (Fig. 3A, C). Further fine mapping of the domain using two different NBS1 truncation mutants (NBS221-402 and NBS402-653) together with mTOR or Rictor showed that only NBS221-402 interacted with mTOR and Rictor (Fig. 3B, D). All the results indicated that the internal domain (a.a. 221-402) of NBS1 interacted with both mTOR and Rictor.

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