FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2002692130> ?p ?o ?g. }

• W2002692130 endingPage "44596" @default.
• W2002692130 startingPage "44593" @default.
• W2002692130 abstract "Mutation in either the TSC1 or TSC2 tumor suppressor gene is responsible for the inherited genetic disease of tuberous sclerosis complex. TSC1 and TSC2 form a physical and functional complex to regulate cell growth. Recently, it has been demonstrated that TSC1·TSC2 functions to inhibit ribosomal S6 kinase and negatively regulate cell size. TSC2 is negatively regulated by Akt phosphorylation. Here, we report that TSC2, but not TSC1, associates with 14-3-3 in vivo. Phosphorylation of Ser1210 in TSC2 is required for its association with 14-3-3. Our data indicate that 14-3-3 association may inhibit the function of TSC2 and represents a possible mechanism of TSC2 regulation. Mutation in either the TSC1 or TSC2 tumor suppressor gene is responsible for the inherited genetic disease of tuberous sclerosis complex. TSC1 and TSC2 form a physical and functional complex to regulate cell growth. Recently, it has been demonstrated that TSC1·TSC2 functions to inhibit ribosomal S6 kinase and negatively regulate cell size. TSC2 is negatively regulated by Akt phosphorylation. Here, we report that TSC2, but not TSC1, associates with 14-3-3 in vivo. Phosphorylation of Ser1210 in TSC2 is required for its association with 14-3-3. Our data indicate that 14-3-3 association may inhibit the function of TSC2 and represents a possible mechanism of TSC2 regulation. Tuberous sclerosis complex (TSC) 1The abbreviations used are: TSC, tuberous sclerosis complex; S6K, ribosomal S6 kinase; mTOR, mammalian target of rapamycin; 4E-BP1, eukaryotic initiation factor 4E-binding protein 1; GST, glutathione S-transferase; HA, hemagglutinin; DMEM, Dulbecco's modified Eagle's medium; FBS, fetal bovine serum; PKA, protein kinase A; PKC, protein kinase C.1The abbreviations used are: TSC, tuberous sclerosis complex; S6K, ribosomal S6 kinase; mTOR, mammalian target of rapamycin; 4E-BP1, eukaryotic initiation factor 4E-binding protein 1; GST, glutathione S-transferase; HA, hemagglutinin; DMEM, Dulbecco's modified Eagle's medium; FBS, fetal bovine serum; PKA, protein kinase A; PKC, protein kinase C. is a relatively common genetic disorder. TSC is caused by mutation in either the TSC1 (hamartin) or TSC2 (tuberin) gene, of which each contributes to ∼50% of the genetic defects (1Young J. Povey S. Mol. Med. Today. 1998; 4: 313-319Google Scholar, 2Crino P.B. Henske E.P. Neurology. 1999; 53: 1384-1390Google Scholar). Studies of TSC patients and animal models support the hypothesis that TSC1 and TSC2 are tumor suppressor genes. Homozygous deletion of either TSC1 or TSC2 in mice produces an embryonic lethal phenotype, suggesting an essential function in development. As predicted, heterozygous deletion of either TSC1 or TSC2 in mice results in a significant increase of carcinomas in many tissues with a 100% incidence of renal carcinomas (3Onda H. Lueck A. Marks P.W. Warren H.B. Kwiatkowski D.J. J. Clin. Invest. 1999; 104: 687-695Google Scholar, 4Kobayashi T. Minowa O. Sugitani Y. Takai S. Mitani H. Kobayashi E. Noda T. Hino O. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 8762-8767Google Scholar). Mutation of TSC1 or TSC2 results in similar phenotypes, suggesting that the two proteins function in the same pathway. Biochemical studies have shown that TSC1 and TSC2 form a stable complex (5van Slegtenhorst M. Nellist M. Nagelkerken B. Cheadle J. Snell R. van den Ouweland A. Reuser A. Sampson J. Halley D. van der Sluijs P. Hum. Mol. Genet. 1998; 7: 1053-1057Google Scholar). Genetic studies inDrosophila have demonstrated that TSC1·TSC2 plays a major negative role in the regulation of cell growth. Mutation of either TSC1 or TSC2 results in a significant increase of cell mass inDrosophila (6Gao X. Pan D. Genes Dev. 2001; 15: 1383-1392Google Scholar, 7Potter C.J. Huang H. Xu T. Cell. 2001; 105: 357-368Google Scholar, 8Tapon N. Ito N. Dickson B.J. Treisman J.E. Hariharan I.K. Cell. 2001; 105: 345-355Google Scholar). Overexpression of either TSC1 or TSC2 inDrosophila produces little phenotype, while co-expression of both TSC1 and TSC2 causes a significant reduction of cell size. Furthermore, genetic epistatic studies indicate that TSC1·TSC2 acts downstream of the insulin receptor (6Gao X. Pan D. Genes Dev. 2001; 15: 1383-1392Google Scholar, 7Potter C.J. Huang H. Xu T. Cell. 2001; 105: 357-368Google Scholar, 8Tapon N. Ito N. Dickson B.J. Treisman J.E. Hariharan I.K. Cell. 2001; 105: 345-355Google Scholar). Recently, we and other groups have demonstrated that TSC1·TSC2 functions to inhibit S6K activation (9Gao X. Zhang Y. Arrazola P. Hino O. Kobayashi T. Yeung R.S. Ru B. Pan D. Nat. Cell Biol. 2002; 4: 699-704Google Scholar, 10Goncharova E.A. Goncharov D.A. Eszterhas A. Hunter D.S. Glassberg M.K. Yeung R.S. Walker C.L. Noonan D. Kwiatkowski D.J. Chou M.M. Panettieri Jr., R.A. Krymskaya V.P. J. Biol. Chem. 2002; 277: 30958-30967Google Scholar, 11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar, 12Porter C.J. Pedraza L.G. Xu T. Nat. Cell Biol. 2002; 4: 658-665Google Scholar, 13Manning B.D. Tee A.R. Logsdon M.N. Blenis J. Cantley L.C. Mol. Cell. 2002; 10: 151-162Google Scholar, 14Kwiatkowski D.J. Zhang H. Bandura J.L. Heiberger K.M. Glogauer M. el-Hashemite N. Onda H. Hum. Mol. Genet. 2002; 11: 525-534Google Scholar). In TSC1−/− or TSC2−/− cells, S6K is highly activated. S6K activation requires phosphorylation of multiple sites. Interestingly, TSC1·TSC2 specifically inhibits the phosphorylation of Thr389, but not phosphorylation of Thr421 and Ser424 in S6K. Thr389is the primary site phosphorylated by mTOR (mammalian target of rapamycin) (15Schmelzle T. Hall M.N. Cell. 2000; 103: 253-262Google Scholar). Furthermore, TSC1·TSC2 also inhibits phosphorylation of 4E-BP1 (eukaryotic initiation factor 4E-binding protein 1), which is also an mTOR target. Both genetic data and biochemical data indicate that TSC1·TSC2 inhibits the function of mTOR (9Gao X. Zhang Y. Arrazola P. Hino O. Kobayashi T. Yeung R.S. Ru B. Pan D. Nat. Cell Biol. 2002; 4: 699-704Google Scholar, 11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar). Several groups, including ours, have shown that TSC2 is directly phosphorylated and inhibited by Akt (11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar, 12Porter C.J. Pedraza L.G. Xu T. Nat. Cell Biol. 2002; 4: 658-665Google Scholar, 13Manning B.D. Tee A.R. Logsdon M.N. Blenis J. Cantley L.C. Mol. Cell. 2002; 10: 151-162Google Scholar). These studies provide an important link between TSC2 and growth factor signaling. In this report, we show that TSC2 interacts with 14-3-3, but not the binding-defective 14-3-3 mutant. This interaction is dependent on the phosphorylation of TSC2. We have identified that Ser1210 of TSC2 is phosphorylated in vivo and is the primary binding site of 14-3-3. In contrast, mutation of all AKT phosphorylation sites in TSC2 had no effect on its interaction with 14-3-3. Overexpression of 14-3-3 enhanced phosphorylation of both S6K and 4E-BP1. Furthermore we demonstrated that interaction between TSC2 and 14-3-3 is also modulated by serum starvation. Anti-phospho-S6K and anti-phospho-4E-BP1 were from Cell Signaling Inc. and anti-TSC2, anti-TSC2 blocking peptide, anti-14-3-3β (K-19), anti-14-3-3β (C-20), anti-14-3-3θ, anti-14-3-3ζ, anti-14-3-3γ were from Santa Cruz Biotechnology. Anti-HA and anti-Myc were from Covance; anti-FLAG and mouse IgG were purchased from Sigma. Rat TSC1 and TSC2 constructs were generously provided by Dr. Y. Xiong. HA-tagged S6K1 and all other DNA constructs including Myc-1433β, Myc-1433β-DN (dominant negative), and FLAG-4E-BP1 were laboratory stocks. Expressions of those plasmids are controlled by the pCMV promoter. Mutant constructs of TSC2 were created by PCR mutagenesis and verified by DNA sequencing. HEK293 cells and Phoenix (retrovirus packaging cells) were seeded and maintained in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS). LExF2 (TSC2−/− cell line) were cultured in DMEM/F-12 containing 10% FBS (16Yeung R.S. Xiao G.H. Jin F. Lee W.C. Testa J.R. Knudson A.G. Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 11413-11416Google Scholar). Transfections were performed using LipofectAMINETM Reagent (Invitrogen) following the manufacturer's instructions. Transiently transfected cells were lysed in lysis buffer (10 mm Tris-HCl, pH 7.5, 100 mm NaCl, 1% Nonidet P-40, 1% Triton X-100, 50 mm NaF, 2 mm EDTA, 1 mmphenylmethylsulfonyl fluoride, 10 μg/ml leupeptin, 10 μg/ml aprotinin) and immunoprecipitated with the indicated antibodies. Immunocomplexes were subjected to SDS-PAGE. HEK293 cells were co-transfected with the indicated plasmids. The serum-starved cells were washed twice with phosphate-free DMEM and incubated with 0.25 mCi/ml [32P]orthophosphate (ICN) for 4 h. HA-tagged TSC2 was immunoprecipitated, resolved by SDS-PAGE, and transferred to a polyvinylidene difluoride membrane. Phosphorylated TSC2 was visualized by autoradiography. Phosphopeptide mapping was performed as described previously (11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar). The TSC2 cDNA was subcloned to the retrovirial vector pPGS-CMV-CITE-Neo. The vectors containing TSC2 were transfected into the Phoenix retrovirus packaging cell line using the calcium phosphate method (Profection Kit, Promega). 48 h post-transfection, retrovirus produced by the Phoenix cells was used for infection of LExF2 cells. LExF2 cells stably expressing TSC2 were selected for and maintained by G418 (300 μg/ml). We have previously found that TSC2 is negatively regulated by AKT phosphorylation (11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar). Akt recognition sequences often overlap with putative 14-3-3 binding sites (17Brunet A. Bonni A. Zigmond M.J. Lin M.Z. Juo P. Hu L.S. Anderson M.J. Arden K.C. Blenis J. Greenberg M.E. Cell. 1999; 96: 857-868Google Scholar, 18Tzivion G. Avruch J. J. Biol. Chem. 2002; 277: 3061-3064Google Scholar). 14-3-3 has been shown to regulate the function of many cellular proteins via a direct association (19Fu H. Subramanian R.R. Masters S.C. Annu. Rev. Pharmacol. Toxicol. 2000; 40: 617-647Google Scholar). We tested whether TSC2 interacts with 14-3-3 in HEK293 cells. Our data showed that 14-3-3β co-precipitated with HA-TSC2 (Fig.1 a). In contrast, a dominant negative 14-3-3, which is unable to bind target proteins (20Thorson J.A. Yu L.W. Hsu A.L. Shih N.Y. Graves P.R. Tanner J.W. Allen P.M. Piwnica-Worms H. Shaw A.S. Mol. Cell. Biol. 1998; 18: 5229-5238Google Scholar), did not associate with TSC1·TSC2 (Fig. 1 a). Reciprocal immunoprecipitation confirmed that TSC2 could be immunoprecipitated with 14-3-3 (Fig. 2 a). 14-3-3 is a family of highly related proteins with numerous isoforms. We found that HEK293 cells express 14-3-3 β, γ, θ, and ζ. Our results showed that TSC2 interacts with all endogenous 14-3-3 isoforms tested present in HEK293 cells (Fig. 1 b). These results suggest that TSC2 interacts with different 14-3-3 isoforms.Figure 2TSC2, but not TSC1, interacts with 14-3-3. a, binding of TSC2 with co-transfected 14-3-3. HEK293 cells were transfected with 100 ng of Myc-TSC1, HA-TSC2, and FLAG-14-3-3β as indicated. The immunoprecipitates were blotted with anti-HA for HA-TSC2, anti- Myc for Myc-TSC1, and anti-FLAG for FLAG-14-3-3. b, binding of TSC2 with endogenous 14-3-3. HEK293 cells were transfected with Myc-TSC1, HA-TSC2, or both plasmids. Co-immunoprecipitation of 14-3-3 was detected by the anti-14-3-3β (K-19) antibody. IP, immunoprecipitation.View Large Image Figure ViewerDownload (PPT) To demonstrate that TSC2 interacts with 14-3-3 under physiological conditions, we immunoprecipitated endogenous TSC2 from 293 cells. Western blot with the anti-14-3-3β (K-19) antibody, which recognizes all 14-3-3 isoforms, indicated that TSC2 associates with 14-3-3 under physiological conditions (Fig. 1 c). Preincubation of the TSC2 antibody with a competing peptide completely eliminated TSC2 and the co-precipitated 14-3-3 (Fig. 1 c). These results demonstrated that TSC2 is associated with 14-3-3 under physiological conditions and suggest that 14-3-3 may play a role in the regulation of TSC2. TSC1 and TSC2 form a physical and functional complex in vivo. To determine whether the TSC1·TSC2 complex, TSC1 or TSC2 alone interacts with 14-3-3, HEK293 cells were transfected with these constructs, and co-immunoprecipitation studies were performed. Immunoprecipitation of 14-3-3 β did not bring down TSC1 alone. Co-precipitation of 14-3-3 and TSC1 was observed only when TSC2 was present in the transfection (Fig. 2 a), indicating that TSC1 alone cannot interact with 14-3-3. In contrast, 14-3-3 co-immunoprecipitated with TSC2 regardless of the presence of TSC1. These results demonstrate that 14-3-3 can interact with TSC2 alone or the TSC1·TSC2 complex. The interaction between transfected TSC2, but not TSC1, and endogenous 14-3-3 further confirmed that 14-3-3 interacts with TSC2 or with the TSC1·TSC2 complex, but not with TSC1 (Fig. 2 b). Interestingly, 14-3-3 interacts with TSC2 stronger than with the TSC1·TSC2 complex (Fig.2). We tested the effect of 14-3-3 on the TSC1·TSC2 complex and found that 14-3-3 had no significant effect on the complex formation between TSC1 and TSC2 (data not shown). Serial deletions of TSC2 were constructed to locate the domain responsible for 14-3-3 interaction. Our results showed that fragments 1–608, 1–1080, 1–1200, and 1321–1765 did not bind 14-3-3, while fragments 1–1320, 1101–1320, 1080–1765 interacted with 14-3-3 at a level similar to the wild type TSC2 (Fig. 3 a). These data demonstrate that the 14-3-3 binding site in TSC2 is localized between residues 1101 and 1320. It has been well established that 14-3-3 binds to phosphorylated residues with a consensus recognition sequence (18Tzivion G. Avruch J. J. Biol. Chem. 2002; 277: 3061-3064Google Scholar). We first tested whether the Akt phosphorylation sites are required for 14-3-3 binding. The TSC2–6A mutant, which has all six predicted Akt phosphorylation sites (residues 939, 1086, 1088, 1378, 1422, 1756) substituted by alanine (11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar), still binds to 14-3-3 at a level no different from wild type TSC2 (Fig.3 b). This observation shows that the 14-3-3 binding site in TSC2 is different from the Akt phosphorylation sites. Sequence analysis by scansite (www.scansite.mit.edu) (21Yaffe M.B. Leparc G.G. Lai J. Obata T. Volinia S. Cantley L.C. Nat. Biotechnol. 2001; 19: 348-353Google Scholar) predicts that TSC2 contains several putative 14-3-3 binding sites. We created single and double mutations by substituting the top four predicted 14-3-3 binding sites (Fig. 3 b). Our data demonstrated that Ser1210 is essential for 14-3-3 binding, while mutations of the other putative sites had no effect on 14-3-3 binding (Fig.3 b). These data are completely consistent with the deletion data that fragment 1101–1320 contains the 14-3-3 binding site. We further mutated Ser1210 in the fragment 1101–1320 and confirmed that Ser1210 is essential for 14-3-3 binding (Fig. 3 b). Therefore, TSC2 utilizes Ser1210 as the primary 14-3-3 binding site. We wanted to test whether the interaction between 14-3-3 and TSC2 requires the phosphorylation of TSC2. GST-TSC2 was expressed and purified from transfected HEK293 cells. Purified GST-TSC2 was treated with λ-phosphatase. Dephosphorylation of GST-TSC2 is evident by an increased electrophoretic mobility of the protein (Fig.4 a). The purified GST-TSC2 was incubated with immunoprecipitated Myc-14-3-3β and the co-precipitation of GST-TSC2 by Myc-14-3-3β was determined. Treatment with phosphatase completely eliminated the interaction between GST-TSC2 and Myc-14-3-3β (Fig. 4 a). To directly demonstrate the phosphorylation status of Ser1210 in TSC2, we performed in vivo 32P labeling and two-dimensional phosphopeptide mapping of TSC2. The TSC2/S1210A mutant eliminated a single phosphopeptide spot depicted by the arrow in Fig. 4 b, while the rest of phosphopeptides were unchanged (Fig. 4 b). These results strongly indicate that Ser1210 is an in vivo phosphorylation site in TSC2. Binding of 14-3-3 may modulate the cellular function of TSC2. We have shown that one of the physiological functions of TSC1·TSC2 is to inhibit S6K activation. In TSC2−/− LExF2 cells, S6K is highly activated. The abilities of wild type and 14-3-3 binding-defective mutant TSC2 to inhibit S6K were tested in the TSC2−/− cells. We observed that both wild type and the 14-3-3 binding-defective TSC2 could inhibit S6K (Fig. 4 c). These data indicate that 14-3-3 binding may not modulate the ability of TSC2 to inhibit S6K. However, the lack of a difference between the wild type and the mutant TSC2 could be due to the fact that the majority of the expressed TSC2 is not phosphorylated on Ser1210, therefore, free of 14-3-3 binding. Our two-dimensional phosphopeptide mapping data also indicates that the majority of TSC2 is not phosphorylated on Ser1210, because the intensity of this phosphopeptide is significantly weaker compared with the Akt phosphorylation site Ser939 (Fig.4 b) (11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar). Sequences surrounding Ser1210 have limited resemblance to PKA and PKC-δ phosphorylation sites. Inhibition of PKA (data not shown) or PKC-δ by rottlerin had no effect on the interaction between TSC2 and 14-3-3 (Fig. 4 d). We also observed that inhibition of the phosphatidylinositol 3-kinase-Akt pathway by wortmannin and the ERK pathway by PD90589 had no effect on the interaction (Fig.4 d), which suggests that the Akt phosphorylation sites are not involved. Interestingly, serum starvation resulted in a visible reduction of association between TSC2 and 14-3-3 (Fig. 4 d). The above data indicate that the complex formation between TSC2 and 14-3-3 may be modulated by cell growth status. To test the effect of 14-3-3 on downstream effectors of TSC2, we examined the phosphorylation of S6K and 4E-BP1. Phosphorylation of these two proteins was inhibited by TSC1·TSC2. We discovered that co-expression of 14-3-3β elevated the Thr389 phosphorylation of S6K (Fig. 4 e). Similarly, expression of 14-3-3β also enhanced the basal phosphorylation of 4E-BP1 (Fig. 4 e). These observations indicate that 14-3-3 may negatively regulate the functions of TSC2. The cellular functions of the TSC1·TSC2 tumor suppressor gene products have just begun to be elucidated. TSC1·TSC2 plays an important role in cell growth regulation and cell size control. Recent studies have demonstrated the TSC2 protein was phosphorylated and inhibited by Akt-dependent phosphorylation (11Inoki K. Li Y. Zhu T. Wu J. Guan K.L. Nat. Cell Biol. 2002; 4: 648-657Google Scholar, 12Porter C.J. Pedraza L.G. Xu T. Nat. Cell Biol. 2002; 4: 658-665Google Scholar, 13Manning B.D. Tee A.R. Logsdon M.N. Blenis J. Cantley L.C. Mol. Cell. 2002; 10: 151-162Google Scholar). In this report, we showed that TSC2 binds to 14-3-3 under physiological conditions. We have mapped a single site, Ser1210, in TSC2 responsible for binding with 14-3-3. The binding of 14-3-3 requires the phosphorylation of Ser1210. During the preparation of this manuscript, Nellist et al. (22Nellist M. Goedbloed M.A. De Winter C. Verhaaf B. Jankie A. Reuser A.J. Van Den Ouweland A.M. Van Der Sluijs P. Halley D.J. J. Biol. Chem. 2002; 277: 39417-39429Google Scholar) also reported that TSC2 interacts with 14-3-3. However, they showed that 14-3-3 binds to multiple sites in TSC2 and concluded that the Akt phosphorylation sites in TSC2 are responsible for 14-3-3 binding (22Nellist M. Goedbloed M.A. De Winter C. Verhaaf B. Jankie A. Reuser A.J. Van Den Ouweland A.M. Van Der Sluijs P. Halley D.J. J. Biol. Chem. 2002; 277: 39417-39429Google Scholar). We have no obvious explanation why the data by Nellist et al. (22Nellist M. Goedbloed M.A. De Winter C. Verhaaf B. Jankie A. Reuser A.J. Van Den Ouweland A.M. Van Der Sluijs P. Halley D.J. J. Biol. Chem. 2002; 277: 39417-39429Google Scholar) are dramatically different from ours. Nevertheless, our data clearly indicate that one major 14-3-3 binding site exists in TSC2, and Akt phosphorylation sites are not responsible for 14-3-3 binding. Overexpression of 14-3-3 results in elevated phosphorylation of S6K and 4E-BP1. In contrast, overexpression of TSC1·TSC2 suppresses the phosphorylation of these two proteins, indicating 14-3-3 and TSC1·TSC2 have opposite effects on S6K activation. Our results indicate that 14-3-3 binds to phosphorylated TSC2 and may suppress its activity. This interpretation is consistent with the fact that the interaction between TSC2 and 14-3-3 is decreased under serum-starved conditions. Serum starvation is predicted to activate TSC2 and suppress cell growth. The dissociation of 14-3-3 may partly contribute to TSC2 activation and S6K inhibition under conditions of serum starvation. However, we cannot exclude the possibility that the effect of 14-3-3 on S6K and 4E-BP1 may not be mediated by TSC2. It has been reported that 14-3-3 may positively modulate the function of TOR in yeast (23Bertram P.G. Zeng C. Thorson J. Shaw A.S. Zheng X.F. Curr. Biol. 1998; 8: 1259-1267Google Scholar). 14-3-3 has also been shown to interact with mTOR (24Mori H. Inoue M. Yano M. Wakabayashi H. Kido H. FEBS Lett. 2000; 467: 61-64Google Scholar). Therefore, 14-3-3 may regulate S6K and 4E-BP1 through multiple targets. Future studies to identify the kinase responsible for phosphorylation of Ser1210 will provide new insights into the mechanism of TSC2 regulation. We thank Tianqing Zhu for technical assistance, Haris Vikis and Jen Aurandt for critical reading of the manuscript, and Yue Xiong for communication of unpublished information." @default.
• W2002692130 created "2016-06-24" @default.
• W2002692130 creator A5023214008 @default.
• W2002692130 creator A5037621688 @default.
• W2002692130 creator A5043534898 @default.
• W2002692130 creator A5072866393 @default.
• W2002692130 date "2002-11-01" @default.
• W2002692130 modified "2023-10-18" @default.
• W2002692130 title "Regulation of TSC2 by 14-3-3 Binding" @default.
• W2002692130 cites W1493799579 @default.
• W2002692130 cites W1968214041 @default.
• W2002692130 cites W1971556742 @default.
• W2002692130 cites W1978625554 @default.
• W2002692130 cites W1978666863 @default.
• W2002692130 cites W1980754624 @default.
• W2002692130 cites W1985046929 @default.
• W2002692130 cites W1987826022 @default.
• W2002692130 cites W1993012047 @default.
• W2002692130 cites W2011282245 @default.
• W2002692130 cites W2021207380 @default.
• W2002692130 cites W2029810183 @default.
• W2002692130 cites W2032685334 @default.
• W2002692130 cites W2048168473 @default.
• W2002692130 cites W2067458366 @default.
• W2002692130 cites W2082459470 @default.
• W2002692130 cites W2094941511 @default.
• W2002692130 cites W2103304405 @default.
• W2002692130 cites W2119979409 @default.
• W2002692130 cites W2122772601 @default.
• W2002692130 cites W2137035181 @default.
• W2002692130 cites W2152973392 @default.
• W2002692130 cites W2155515046 @default.
• W2002692130 cites W2170936754 @default.
• W2002692130 doi "https://doi.org/10.1074/jbc.c200510200" @default.
• W2002692130 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/12364343" @default.
• W2002692130 hasPublicationYear "2002" @default.
• W2002692130 type Work @default.
• W2002692130 sameAs 2002692130 @default.
• W2002692130 citedByCount "105" @default.
• W2002692130 countsByYear W20026921302012 @default.
• W2002692130 countsByYear W20026921302013 @default.
• W2002692130 countsByYear W20026921302014 @default.
• W2002692130 countsByYear W20026921302015 @default.
• W2002692130 countsByYear W20026921302016 @default.
• W2002692130 countsByYear W20026921302017 @default.
• W2002692130 countsByYear W20026921302018 @default.
• W2002692130 countsByYear W20026921302019 @default.
• W2002692130 countsByYear W20026921302020 @default.
• W2002692130 countsByYear W20026921302021 @default.
• W2002692130 countsByYear W20026921302022 @default.
• W2002692130 countsByYear W20026921302023 @default.
• W2002692130 crossrefType "journal-article" @default.
• W2002692130 hasAuthorship W2002692130A5023214008 @default.
• W2002692130 hasAuthorship W2002692130A5037621688 @default.
• W2002692130 hasAuthorship W2002692130A5043534898 @default.
• W2002692130 hasAuthorship W2002692130A5072866393 @default.
• W2002692130 hasBestOaLocation W20026921301 @default.
• W2002692130 hasConcept C185592680 @default.
• W2002692130 hasConcept C55493867 @default.
• W2002692130 hasConcept C86803240 @default.
• W2002692130 hasConcept C95444343 @default.
• W2002692130 hasConceptScore W2002692130C185592680 @default.
• W2002692130 hasConceptScore W2002692130C55493867 @default.
• W2002692130 hasConceptScore W2002692130C86803240 @default.
• W2002692130 hasConceptScore W2002692130C95444343 @default.
• W2002692130 hasIssue "47" @default.
• W2002692130 hasLocation W20026921301 @default.
• W2002692130 hasLocation W20026921302 @default.
• W2002692130 hasOpenAccess W2002692130 @default.
• W2002692130 hasPrimaryLocation W20026921301 @default.
• W2002692130 hasRelatedWork W1531601525 @default.
• W2002692130 hasRelatedWork W2319480705 @default.
• W2002692130 hasRelatedWork W2384464875 @default.
• W2002692130 hasRelatedWork W2398689458 @default.
• W2002692130 hasRelatedWork W2606230654 @default.
• W2002692130 hasRelatedWork W2607424097 @default.
• W2002692130 hasRelatedWork W2748952813 @default.
• W2002692130 hasRelatedWork W2899084033 @default.
• W2002692130 hasRelatedWork W2948807893 @default.
• W2002692130 hasRelatedWork W2778153218 @default.
• W2002692130 hasVolume "277" @default.
• W2002692130 isParatext "false" @default.
• W2002692130 isRetracted "false" @default.
• W2002692130 magId "2002692130" @default.
• W2002692130 workType "article" @default.