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Published Erratum
. 2021 Aug 17;118(33):e2112628118.
doi: 10.1073/pnas.2112628118.

Correction for Tee et al., Tuberous sclerosis complex-1 and -2 gene products function together to inhibit mammalian target of rapamycin (mTOR)-mediated downstream signaling

No authors listed
Published Erratum

Correction for Tee et al., Tuberous sclerosis complex-1 and -2 gene products function together to inhibit mammalian target of rapamycin (mTOR)-mediated downstream signaling

No authors listed. Proc Natl Acad Sci U S A. .
No abstract available

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Figures

Fig. 1.
Fig. 1.
Hamartin and tuberin inhibits PI3K-dependent 4E-BP1 phosphorylation. HEK293E cells coexpressing Flag-tagged hamartin (Ham) and tuberin (Tub), where indicated, with HA-tagged 4E-BP1 were serum-starved and pretreated with 20 nM rapamycin (rap) for 30 min before being stimulated with insulin (100 nM) or PMA (100 ng/mL) for 30 min, where indicated. (A) The levels and Thr-308 phosphorylation of Akt and the levels and phosphorylation of the MAPK isoforms (p44 and p42) were determined. (B) Hamartin and tuberin protein levels were accessed from the cell lysates (Sol) and the insoluble fraction (Non-Sol) as described in Materials and Methods by using the anti-Flag antibody. Thr-1462 tuberin phosphorylation was analyzed by using a tuberin Thr-1462 phospho-specific antibody. (C) The phosphorylation of exogenous 4E-BP1 was determined with an anti-HA antibody and phospho-specific antibodies for 4E-BP1 at Thr-37 and/or 46, Ser-65, and Thr-70, as indicated. The α-, β-, and γ-species of 4E-BP1 are labeled accordingly. (D) Cell extracts were subjected to affinity chromatography on m7GTP-Sepharose, as described in Materials and Methods. The levels of eIF4E and exogenous 4E-BP1 that was copurified were determined.
Fig. 2.
Fig. 2.
Hamartin and tuberin inhibit 4E-BP1 phosphorylation and S6K1 activity within proliferating cells. U20S cells overexpressing 4E-BP1 (A) or S6K1 (B) with or without hamartin (Ham) and tuberin (Tub), where indicated, were grown in serum and treated with 20 nM of rapamycin (Rap) for 30 min, as indicated. Hamartin, tuberin, and MAPK isoform (as a loading control) protein levels are shown. The α-, β-, and γ-species of 4E-BP1 are labeled accordingly. S6K1 kinase assays were carried out as described in Materials and Methods. The total levels of S6K1 are shown. Incorporation of 32P label into GST-S6 was assessed, and an autoradiograph of the gel is presented. The ratios of 32P label incorporated into GST-S6 were normalized against the empty vector (pRK7). The data presented are representative of at least three experiments. (C) HEK293E cells overexpressing 4E-BP1 with or without hamartin (Ham), tuberin (Tub), and the tuberin K599M mutant [Tub(K599M)], where indicated, were serum-starved and then stimulated with 100 nM insulin for 30 min, where indicated. Hamartin and tuberin expression and the extent of phosphorylation of 4E-BP1 was analyzed as for Fig. 1C.
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