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A phospho-proteomic screen identifies novel S6K1 and mTORC1 substrates revealing additional complexity in the signaling network regulating cell growth

机译:磷酸化蛋白质组学筛选可识别新型S6K1和mTORC1底物,揭示了调控细胞生长的信号网络中的其他复杂性

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摘要

S6K1, a critical downstream substrate of mTORC1, has been implicated in regulating protein synthesis and a variety of processes that impinge upon cell growth and proliferation. While the role of the cytoplasmic p70~(S6K1) isoform in the regulation of translation has been intensively studied, the targets and function of the nuclear p85~(S6K1) isoform remain unclear. Therefore, we carried out a phospho-proteomic screen to identify novel p85~(S6K1) substrates. Four novel putative p85~(S6K1) substrates, GRP75, CCTβ, PGK1 and RACK1, and two mTORC1 substrates, ANXA4 and PSMA6 were identified, with diverse roles in chaperone function, ribosome maturation, metabolism, vesicle trafficking and the proteasome, respectively. The chaperonin subunit CCTβ was further investigated and the site of phosphorylation mapped to serine 260, a site located in the chaperonin apical domain. Consistent with this domain being involved in folding substrate interactions, we found that phosphorylation of serine 260 modulates chaperonin folding activity.
机译:S6K1是mTORC1的关键下游底物,已参与调节蛋白质的合成以及影响细胞生长和增殖的各种过程。虽然已经深入研究了胞质p70〜(S6K1)同工型在翻译调控中的作用,但仍不清楚核p85〜(S6K1)同工型的靶标和功能。因此,我们进行了磷酸化蛋白质组学筛选,以鉴定新型的p85〜(S6K1)底物。鉴定出四种新颖的假定的p85〜(S6K1)底物GRP75,CCTβ,PGK1和RACK1,以及两种mTORC1底物ANXA4和PSMA6,它们分别在分子伴侣功能,核糖体成熟,代谢,囊泡运输和蛋白酶体中具有不同的作用。进一步研究了伴侣蛋白亚基CCTβ,并将磷酸化位点映射到丝氨酸260,丝氨酸260位于伴侣蛋白心尖区。与该结构域参与折叠底物相互作用一致,我们发现丝氨酸260的磷酸化调节伴侣蛋白折叠活性。

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