title: Tel2 structure and function in the Hsp90-dependent maturation of mTOR and ATR complexes.
abstract: We reported previously that the stability of all mammalian phosphatidylinositol 3-kinase-related protein kinases (PIKKs) depends on their interaction with Tel2, the ortholog of yeast Tel2 and Caenorhabditis elegans Clk-2. Here we provide evidence that Tel2 acts with Hsp90 in the maturation of PIKK complexes. Quantitative immunoblotting showed that the abundance of Tel2 is low compared with the PIKKs, and Tel2 preferentially bound newly synthesized ATM, ATR, mTOR, and DNA-PKcs. Tel2 complexes contained, in addition to Tti1-Tti2, the Hsp90 chaperone, and inhibition of Hsp90 interfered with the interaction of Tel2 with the PIKKs. Analysis of in vivo labeled nascent protein complexes showed that Tel2 and Hsp90 mediate the formation of the mTOR TORC1 and TORC2 complexes and the association of ATR with ATRIP. The structure of yeast Tel2, reported here, shows that Tel2 consists of HEAT-like helical repeats that assemble into two separate α-solenoids. Through mutagenesis, we identify a surface patch of conserved residues involved in binding to the Tti1-Tti2 complex in vitro. In vivo, mutation of this conserved patch affects cell growth, levels of PIKKs, and ATM/ATR-mediated checkpoint signaling, highlighting the importance of Tti1-Tti2 binding to the function of Tel2. Taken together, our data suggest that the Tel2-Tti1-Tti2 complex is a PIKK-specific cochaperone for Hsp90.
authors.affiliation: Laboratory for Cell Biology and Genetics, The Rockefeller University, New York, New York 10065, USA.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takai
authors.forename: Yihu
authors.initials: Y
authors.lastname: Xie
authors.forename: Titia
authors.initials: T
authors.lastname: de Lange
authors.forename: Nikola P
authors.initials: NP
authors.lastname: Pavletich
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000818:Animals; D002478:Cells, Cultured; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics*; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D058570:TOR Serine-Threonine Kinases; D034501:Telomere-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D018841:HSP90 Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D029701:Saccharomyces cerevisiae Proteins; C444420:Tel2 protein, S cerevisiae; C528081:Tel2 protein, mouse; D034501:Telomere-Binding Proteins; C546842:MTOR protein, human; C546843:mTOR protein, mouse; D017346:Protein Serine-Threonine Kinases; D058570:TOR Serine-Threonine Kinases; D000074183:ATPases Associated with Diverse Cellular Activities; D004265:DNA Helicases; C578681:RUVBL1 protein, mouse
references.citation: Oncogene. 2003 Jun 19;22(25):3867-74
references.pmid: 12813460
references.citation: Nat Rev Mol Cell Biol. 2008 Aug;9(8):616-27
references.pmid: 18594563
references.citation: Biochem Cell Biol. 2010 Feb;88(1):29-40
references.pmid: 20130677
references.citation: Genes Dev. 2008 Apr 1;22(7):854-9
references.pmid: 18334620
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Bioinformatics. 2008 Dec 1;24(23):2780-1
references.pmid: 18818215
references.citation: Cancer Cell. 2007 Jul;12(1):9-22
references.pmid: 17613433
references.citation: J Biol Chem. 2007 Feb 23;282(8):5346-55
references.pmid: 17189249
references.citation: J Neurosci. 2003 Mar 15;23(6):2314-22
references.pmid: 12657690
references.citation: Acta Crystallogr A. 2008 Jan;64(Pt 1):112-22
references.pmid: 18156677
references.citation: Oncogene. 2007 Aug 13;26(37):5358-72
references.pmid: 17694078
references.citation: Nat Rev Cancer. 2008 Jul;8(7):491-5
references.pmid: 18511936
references.citation: Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
references.pmid: 15299374
references.citation: J Biol Chem. 2010 Jun 25;285(26):20109-16
references.pmid: 20427287
references.citation: Nat Rev Cancer. 2005 Oct;5(10):761-72
references.pmid: 16175177
references.citation: J Struct Biol. 2001 May-Jun;134(2-3):117-31
references.pmid: 11551174
references.citation: Mol Cell. 2009 Jun 12;34(5):521-33
references.pmid: 19524533
references.citation: DNA Repair (Amst). 2004 Aug-Sep;3(8-9):883-7
references.pmid: 15279773
references.citation: J Mol Biol. 2001 May 25;309(1):1-18
references.pmid: 11491282
references.citation: Science. 2003 Jun 6;300(5625):1542-8
references.pmid: 12791985
references.citation: J Cell Biol. 2001 Jul 23;154(2):267-73
references.pmid: 11470816
references.citation: Cell. 2004 Jul 9;118(1):9-17
references.pmid: 15242640
references.citation: Cell. 2002 Jul 26;110(2):163-75
references.pmid: 12150925
references.citation: Acta Crystallogr A. 1991 Mar 1;47 ( Pt 2):110-9
references.pmid: 2025413
references.citation: Mol Cell. 2007 Aug 3;27(3):509-16
references.pmid: 17679098
references.citation: J Biochem. 2006 Jan;139(1):129-35
references.pmid: 16428328
references.citation: Nat Rev Mol Cell Biol. 2008 Apr;9(4):297-308
references.pmid: 18285803
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: Nat Rev Cancer. 2003 Mar;3(3):155-68
references.pmid: 12612651
references.citation: Sci Signal. 2010 Apr 06;3(116):ra27
references.pmid: 20371770
references.citation: Genome Biol. 2008;9(11):R167
references.pmid: 19040720
references.citation: Mol Cell Proteomics. 2010 Feb;9(2):255-70
references.pmid: 19875381
references.citation: Mol Biol Cell. 2005 May;16(5):2372-81
references.pmid: 15743907
references.citation: Mol Cell. 2007 Dec 14;28(5):739-45
references.pmid: 18082599
references.citation: Mol Cell. 2007 Jul 20;27(2):262-74
references.pmid: 17643375
references.citation: Cell Mol Life Sci. 2010 Mar;67(5):677-700
references.pmid: 19859661
references.citation: J Cell Biol. 2008 Feb 11;180(3):579-95
references.pmid: 18268104
references.citation: J Biol Chem. 2008 Jul 4;283(27):18473-7
references.pmid: 18442971
references.citation: J Proteome Res. 2007 May;6(5):1963-73
references.pmid: 17348703
references.citation: J Cell Sci. 2009 Oct 15;122(Pt 20):3589-94
references.pmid: 19812304
references.citation: Nature. 2010 Jan 7;463(7277):118-21
references.pmid: 20023628
references.citation: Mol Cell. 2006 Apr 21;22(2):159-68
references.pmid: 16603397
references.citation: Proc Natl Acad Sci U S A. 1986 Mar;83(5):1398-402
references.pmid: 3513174
references.citation: Science. 1996 May 17;272(5264):1010-3
references.pmid: 8638122
references.citation: Cell. 2007 Dec 28;131(7):1248-59
references.pmid: 18160036
references.citation: Nat Rev Cancer. 2008 Sep;8(9):665-9
references.pmid: 18633356
references.citation: EMBO J. 2010 Mar 17;29(6):1045-54
references.pmid: 20150893
references.citation: Genes Cells. 2007 Dec;12(12):1357-70
references.pmid: 18076573
references.citation: Genetics. 1982 Oct;102(2):159-78
references.pmid: 7152245
references.citation: Nat Rev Mol Cell Biol. 2009 May;10(5):307-18
references.pmid: 19339977