Limits...
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.

Show MeSH

Related in: MedlinePlus

Mapping of the domain in mTOR or Rictor interacting with NBS1.A & B. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of mTOR and NBS1. Whole cell extracts were shown in A. C & D. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of Rictor and NBS1. Whole cell extracts were shown in C.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3675082&req=5

pone-0065586-g002: Mapping of the domain in mTOR or Rictor interacting with NBS1.A & B. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of mTOR and NBS1. Whole cell extracts were shown in A. C & D. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of Rictor and NBS1. Whole cell extracts were shown in C.

Mentions: In order to map the domain in mTOR or Rictor that interacts with NBS1, different truncation mutants of mTOR or Rictor were generated. Different truncation mutants of mTOR (mTOR1-651, mTOR640-1418, and mTOR1401-2549) were co-expressed with NBS1 in 293T cells followed by co-immunoprecipitation. The results showed that only mTOR1-651 interacted with NBS1 (Fig. 2A–B). The full length and different truncation mutants of Rictor (Rictor1-1708, Rictor1-789, and Rictor664-1708) were co-expressed with NBS1 in 293T cells followed by co-immunoprecipitation. Co-immunoprecipitation experiments showed that the domain 1-789 a.a. of Rictor interacted with NBS1 (Fig. 2C–D). These results indicated that the N-terminal domains (1-651 a.a. domain of mTOR and 1-789 a.a. domain of Rictor) of both proteins interacted with NBS1.


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 mTOR or Rictor interacting with NBS1.A & B. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of mTOR and NBS1. Whole cell extracts were shown in A. C & D. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of Rictor and NBS1. Whole cell extracts were shown in C.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0065586-g002: Mapping of the domain in mTOR or Rictor interacting with NBS1.A & B. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of mTOR and NBS1. Whole cell extracts were shown in A. C & D. Co-immunoprecipitation assays showed the interaction between the N-terminal domain of Rictor and NBS1. Whole cell extracts were shown in C.
Mentions: In order to map the domain in mTOR or Rictor that interacts with NBS1, different truncation mutants of mTOR or Rictor were generated. Different truncation mutants of mTOR (mTOR1-651, mTOR640-1418, and mTOR1401-2549) were co-expressed with NBS1 in 293T cells followed by co-immunoprecipitation. The results showed that only mTOR1-651 interacted with NBS1 (Fig. 2A–B). The full length and different truncation mutants of Rictor (Rictor1-1708, Rictor1-789, and Rictor664-1708) were co-expressed with NBS1 in 293T cells followed by co-immunoprecipitation. Co-immunoprecipitation experiments showed that the domain 1-789 a.a. of Rictor interacted with NBS1 (Fig. 2C–D). These results indicated that the N-terminal domains (1-651 a.a. domain of mTOR and 1-789 a.a. domain of Rictor) of both proteins interacted with NBS1.

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