p90 Ribosomal S6 Kinase and p70 Ribosomal S6 Kinase Link Phosphorylation of the Eukaryotic Chaperonin Containing TCP-1 to Growth Factor, Insulin, and Nutrient Signaling

被引:77
作者
Abe, Yuki [1 ]
Yoon, Sang-Oh [1 ]
Kubota, Kazuishi [1 ]
Mendoza, Michelle C. [1 ]
Gygi, Steven P. [1 ]
Blenis, John [1 ]
机构
[1] Harvard Univ, Sch Med, Dept Cell Biol, Boston, MA 02115 USA
基金
美国国家卫生研究院;
关键词
TUMOR-SUPPRESSOR COMPLEX; IN-VIVO; MOLECULAR CHAPERONES; CYTOPLASMIC CHAPERONIN; PROTEIN-SYNTHESIS; NASCENT CHAIN; AKT SUBSTRATE; IDENTIFICATION; RSK; ACTIN;
D O I
10.1074/jbc.M900097200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Chaperonin containing TCP-1 (CCT) is a large multisubunit complex that mediates protein folding in eukaryotic cells. CCT participates in the folding of newly synthesized polypeptides, including actin, tubulin, and several cell cycle regulators; therefore, CCT plays an important role in cytoskeletal organization and cell division. Here we identify the chaperonin CCT as a novel physiological substrate for p90 ribosomal S6 kinase (RSK) and p70 ribosomal S6 kinase (S6K). RSK phosphorylates the beta subunit of CCT in response to tumor promoters or growth factors that activate the Ras-mitogen-activated protein kinase (MAPK) pathway. CCT beta Ser-260 was identified as the RSK site by mass spectrometry and confirmed by site-directed mutagenesis. RSK-dependent Ser-260 phosphorylation was sensitive to the MEK inhibitor UO126 and the RSK inhibitor BID-1870. Insulin weakly activates RSK but strongly activates the phosphoinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) pathway and utilizes S6K to regulate CCT beta phosphorylation. Thus, the Ras-MAPK and PI3K-mTOR pathways converge on CCT beta Ser-260 phosphorylation in response to multiple agonists in various mammalian cells. We also show that RNA interference-mediated knockdown of endogenous CCT beta causes impaired cell proliferation that can be rescued with ectopically expressed murine CCT beta wild-type or phosphomimetic mutant S260D, but not the phosphorylation-deficient mutant S260A. Although the molecular mechanism of CCT beta regulation remains unclear, our findings demonstrate a link between oncogene and growth factor signaling and chaperonin CCT-mediated cellular activities.
引用
收藏
页码:14939 / 14948
页数:10
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