The role of structural disorder in cell cycle regulation, related clinical proteomics, disease development and drug targeting

Expert Review of Proteomics

Volumn 12, Issue 3, 2015, Pages 221-233

  Tantos, Agnes ^a   Kalmar, Lajos ^b   Tompa, Peter ^a,b  

^a INSTITUTE OF EXPERIMENTAL MEDICINE   (Hungary)

^b VRIJE UNIVERSITEIT BRUSSEL   (Belgium)

Author keywords

cancer; cell cycle; checkpoint; post translational modification; protein disorder; signal transduction

Indexed keywords

CELL CYCLE PROTEIN;

CELL CYCLE; CELL CYCLE CHECKPOINT; CELL CYCLE REGULATION; CELL MEMBRANE; CYTOLOGY; DISEASE COURSE; DRUG TARGETING; EFFECTOR CELL; EUKARYOTE; GENETIC TRANSCRIPTION; HUMAN; MOLECULAR BIOLOGY; NONHUMAN; PHYSICAL DISEASE; PROTEOMICS; REVIEW; ANIMAL; CELL STRUCTURE; CLINICAL MEDICINE; DISEASES; DRUG DELIVERY SYSTEM; METABOLISM; SIGNAL TRANSDUCTION;

EUKARYOTA;

ANIMALS; CELL CYCLE CHECKPOINTS; CELLULAR STRUCTURES; CLINICAL MEDICINE; DISEASE; DRUG DELIVERY SYSTEMS; HUMANS; PROTEOMICS; SIGNAL TRANSDUCTION;

EID: 84929575458     PISSN: 14789450     EISSN: 17448387     Source Type: Journal    
DOI: 10.1586/14789450.2015.1042866     Document Type: Review

References (115)

1. Wright PE, Dyson HJ. Intrinsically disordered proteins in cellular signalling and regulation. Nat Rev Mol Cell Biol 2015;16: 18-29
2. Yoon MK, Mitrea DM, Ou L, Kriwacki RW. Cell cycle regulation by the intrinsically disordered proteins p21 and p27. Biochem Soc Trans 2012;40:981-8
3. Habchi J, Tompa P, Longhi S, Uversky VN. Introducing protein intrinsic disorder. Chem Rev 2014;114:6561-88 .. A comprehensive collection of our current knowledge about the function and evolution of intrinsically disordered proteins. A historic overview of the evolution of the field as well as a description of methods for studying intrinsically disordered proteins (IDPs) is also given.
4. van der Lee R, Buljan M, Lang B, et al. Classification of intrinsically disordered regions and proteins. Chem Rev 2014;114: 6589-631
5. Iakoucheva LM, Brown CJ, Lawson JD, et al. Intrinsic disorder in cell-signaling and cancer-associated proteins. J Mol Biol 2002;323:573-84
6. Tompa P. Unstructural biology coming of age. Curr Opin Struct Biol 2011;21:419-25
7. Funk JO. Cell Cycle Checkpoint Genes and Cancer. Encyclopedia of life sciences 2005
8. Resnitzky D, Hengst L, Reed SI. Cyclin A-associated kinase activity is rate limiting for entrance into S phase and is negatively regulated in G1 by p27Kip1. Mol Cell Biol 1995;15:4347-52
9. Lukas J, Herzinger T, Hansen K, et al. Cyclin E-induced S phase without activation of the pRb/E2F pathway. Genes Dev 1997;11:1479-92
10. Weber HO, Samuel T, Rauch P, Funk JO. Human p14(ARF)-mediated cell cycle arrest strictly depends on intact p53 signaling pathways. Oncogene 2002;21:3207-12
11. Chan TA, Hermeking H, Lengauer C, et al. 14-3-3Sigma is required to prevent mitotic catastrophe after DNA damage. Nature 1999;401:616-20
12. Vermeulen K, Van Bockstaele DR, Berneman ZN. The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif 2003;36: 131-49 .. This review focuses on mechanisms that ensure correct cell division, that is, regulation of cyclin-dependent kinase (CDKs) by cyclins, CDK inhibitors and phosphorylating events and the quality checkpoints activated after DNA damage. It provides an overview of deregulation of the cell cycle in cancer.
13. Willis N, Rhind N. Regulation of DNA replication by the S-phase DNA damage checkpoint. Cell Div 2009;4:13
14. Falck J, Mailand N, Syljuasen RG, et al. The ATM-Chk2-Cdc25A checkpoint pathway guards against radioresistant DNA synthesis. Nature 2001;410:842-7
15. Mailand N, Falck J, Lukas C, et al. Rapid destruction of human Cdc25A in response to DNA damage. Science 2000;288:1425-9
16. Sorensen CS, Syljuasen RG. Safeguarding genome integrity: the checkpoint kinases ATR, CHK1 and WEE1 restrain CDK activity during normal DNA replication. Nucleic Acids Res 2012;40:477-86
17. Unsal-Kacmaz K, Chastain PD, Qu PP, et al. The human Tim/Tipin complex coordinates an Intra-S checkpoint response to UV that slows replication fork displacement. Mol Cell Biol 2007;27: 3131-42
18. de Klein A, Muijtjens M, van Os R, et al. Targeted disruption of the cell-cycle checkpoint gene ATR leads to early embryonic lethality in mice. Curr Biol 2000;10:479-82
19. Tominaga Y, Li C, Wang RH, Deng CX. Murine Wee1 plays a critical role in cell cycle regulation and pre-implantation stages of embryonic development. Int J Biol Sci 2006;2:161-70
20. Lee S, Bolanos-Garcia VM. The dynamics of signal amplification by macromolecular assemblies for the control of chromosome segregation. Front Physiol 2014;5:368 . The highly dynamic nature of multiprotein complexes that control chromosome segregation is discussed. The current structural understanding of the communication between the spindle assembly checkpoint (SAC) and the kinetochore is also discussed along with the mode of transient interactions can regulating the assembly and disassembly of the SAC as well as the challenges and opportunities for the definition and the manipulation of the flow of information in SAC signaling.
21. Tompa P, Varadi M. Predicting the predictive power of IDP ensembles. Structure 2014;22:177-8
22. Varadi M, Kosol S, Lebrun P, et al. pE-DB: a database of structural ensembles of intrinsically disordered and of unfolded proteins. Nucleic Acids Res 2014;42: D326-35
23. Tompa P. The interplay between structure and function in intrinsically unstructured proteins. FEBS Lett 2005;579:3346-54
24. Besson A, Dowdy SF, Roberts JM. CDK inhibitors: cell cycle regulators and beyond. Dev Cell 2008;14:159-69 . The complex functions of the cyclin-CDK inhibitors of the Cip/Kip family are discussed along with the regulation network that modulates their functions by altering their subcellular localization, protein-protein interactions and stability.
25. Cheng M, Olivier P, Diehl JA, et al. The p21(Cip1) and p27(Kip1) CDK 'inhibitors' are essential activators of cyclin D-dependent kinases in murine fibroblasts. EMBO J 1999;18:1571-83
26. LaBaer J, Garrett MD, Stevenson LF, et al. New functional activities for the p21 family of CDK inhibitors. Genes Dev 1997;11: 847-62
27. Wang Y, Fisher JC, Mathew R, et al. Intrinsic disorder mediates the diverse regulatory functions of the Cdk inhibitor p21. Nat Chem Biol 2011;7:214-21
28. Lacy ER, Wang Y, Post J, et al. Molecular basis for the specificity of p27 toward cyclin-dependent kinases that regulate cell division. J Mol Biol 2005;349:764-73
29. Shoemaker BA, Portman JJ, Wolynes PG. Speeding molecular recognition by using the folding funnel: the fly-casting mechanism. Proc Natl Acad Sci USA 2000;97:8868-73
30. Quelle DE, Zindy F, Ashmun RA, Sherr CJ. Alternative reading frames of the INK4a tumor suppressor gene encode two unrelated proteins capable of inducing cell cycle arrest. Cell 1995;83:993-1000
31. Bothner B, Lewis WS, DiGiammarino EL, et al. Defining the molecular basis of Arf and Hdm2 interactions. J Mol Biol 2001;314:263-77
32. Bates S, Phillips AC, Clark PA, et al. p14ARF links the tumour suppressors RB and p53. Nature 1998;395:124-5
33. Zindy F, Eischen CM, Randle DH, et al. Myc signaling via the ARF tumor suppressor regulates p53-dependent apoptosis and immortalization. Genes Dev 1998;12:2424-33
34. Saha T, Kar R K, Sa G. Structural and sequential context of p53: A review of experimental and theoretical evidence. Prog Biophys Mol Biol 2015. [Epub ahead of print]
35. Dawson R, Muller L, Dehner A, et al. The N-terminal domain of p53 is natively unfolded. J Mol Biol 2003;332:1131-41
36. Kussie PH, Gorina S, Marechal V, et al. Structure of the MDM2 oncoprotein bound to the p53 tumor suppressor transactivation domain. Science 1996;274:948-53
37. Lukman S, Lane DP, Verma CS. Mapping the structural and dynamical features of multiple p53 DNA binding domains: insights into loop 1 intrinsic dynamics. PLoS One 2013;8:e80221
38. Hegyi H, Schad E, Tompa P. Structural disorder promotes assembly of protein complexes. BMC Struct Biol 2007;7:65
39. Ghongane P, Kapanidou M, Asghar A, et al. The dynamic protein Knl1 - a kinetochore rendezvous. J Cell Sci 2014;127:3415-23
40. Caldas GV, DeLuca KF, DeLuca JG. KNL1 facilitates phosphorylation of outer kinetochore proteins by promoting Aurora B kinase activity. J Cell Biol 2013;203: 957-69
41. Kiyomitsu T, Obuse C, Yanagida M. Human Blinkin/AF15q14 is required for chromosome alignment and the mitotic checkpoint through direct interaction with Bub1 and BubR1. Dev Cell 2007;13: 663-76
42. Petrovic A, Pasqualato S, Dube P, et al. The MIS12 complex is a protein interaction hub for outer kinetochore assembly. J Cell Biol 2010;190:835-52
43. Kops GJ, Kim Y, Weaver BA, et al. ZW10 links mitotic checkpoint signaling to the structural kinetochore. J Cell Biol 2005;169:49-60
44. Liu D, Vleugel M, Backer CB, et al. Regulated targeting of protein phosphatase 1 to the outer kinetochore by KNL1 opposes Aurora B kinase. J Cell Biol 2010;188:809-20
45. Espeut J, Cheerambathur DK, Krenning L, et al. Microtubule binding by KNL-1 contributes to spindle checkpoint silencing at the kinetochore. J Cell Biol 2012;196:469-82
46. London N, Ceto S, Ranish JA, Biggins S. Phosphoregulation of Spc105 by Mps1 and PP1 regulates Bub1 localization to kinetochores. Curr Biol 2012;22:900-6
47. Kruse T, Zhang G, Larsen MS, et al. Direct binding between BubR1 and B56-PP2A phosphatase complexes regulate mitotic progression. J Cell Sci 2013;126: 1086-92
48. Petrovic A, Mosalaganti S, Keller J, et al. Modular assembly of RWD domains on the Mis12 complex underlies outer kinetochore organization. Mol Cell 2014;53:591-605
49. Coster G, Goldberg M. The cellular response to DNA damage: a focus on MDC1 and its interacting proteins. Nucleus 2010;1:166-78
50. Stewart GS, Wang B, Bignell CR, et al. MDC1 is a mediator of the mammalian DNA damage checkpoint. Nature 2003;421:961-6
51. Lou Z, Chen BP, Asaithamby A, et al. MDC1 regulates DNA-PK autophosphorylation in response to DNA damage. J Biol Chem 2004;279: 46359-62
52. Eliezer Y, Argaman L, Rhie A, et al. The direct interaction between 53BP1 and MDC1 is required for the recruitment of 53BP1 to sites of damage. J Biol Chem 2009;284:426-35
53. Townsend K, Mason H, Blackford AN, et al. Mediator of DNA damage checkpoint 1 (MDC1) regulates mitotic progression. J Biol Chem 2009;284:33939-48
54. Mark WY, Liao JC, Lu Y, et al. Characterization of segments from the central region of BRCA1: an intrinsically disordered scaffold for multiple protein-protein and protein-DNA interactions? J Mol Biol 2005;345:275-87
55. Brzovic PS, Keeffe JR, Nishikawa H, et al. Binding and recognition in the assembly of an active BRCA1/BARD1 ubiquitin-ligase complex. Proc Natl Acad Sci U S A 2003;100:5646-51
56. Williams RS, Green R, Glover JN. Crystal structure of the BRCT repeat region from the breast cancer-associated protein BRCA1. Nat Struct Biol 2001;8:838-42
57. Savage KI, Harkin DP. BRCA1, a 'complex' protein involved in the maintenance of genomic stability. FEBS J 2015;282:630-46
58. Xie J, Peng M, Guillemette S, et al. FANCJ/BACH1 acetylation at lysine 1249 regulates the DNA damage response. PLoS Genet 2012;8:e1002786
59. Greenberg RA, Sobhian B, Pathania S, et al. Multifactorial contributions to an acute DNA damage response by BRCA1/BARD1- containing complexes. Genes Dev 2006;20: 34-46
60. Zhang F, Ma J, Wu J, et al. PALB2 links BRCA1 and BRCA2 in the DNA-damage response. Curr Biol 2009;19:524-9
61. Iakoucheva LM, Radivojac P, Brown CJ, et al. The importance of intrinsic disorder for protein phosphorylation. Nucleic Acids Res 2004;32:1037-49
62. Tyanova S, Cox J, Olsen J, et al. Phosphorylation variation during the cell cycle scales with structural propensities of proteins. PLoS Comput Biol 2013;9: e1002842 .. The authors combine dynamic properties of the phosphoproteome with protein structural features, investigating how the variation of the amount of phosphorylation during the progression of cell cycle correlates with the protein structure in the vicinity of the modified site. The authors found two distinct phosphorylation site groups: intrinsically disordered regions tend to contain sites with dynamically varying levels, whereas regions with predominantly regular secondary structures retain more constant phosphorylation levels. The results suggest that the structural organization of the region in which a phosphorylation site resides may serve as an additional control mechanism.
63. Galea CA, Nourse A, Wang Y, et al. Role of intrinsic flexibility in signal transduction mediated by the cell cycle regulator, p27 Kip1. J Mol Biol 2008;376:827-38
64. Nash P, Tang X, Orlicky S, et al. Multisite phosphorylation of a CDK inhibitor sets a threshold for the onset of DNA replication. Nature 2001;414:514-21
65. Schweiger R, Linial M. Cooperativity within proximal phosphorylation sites is revealed from large-scale proteomics data. Biol Direct 2010;5:6
66. Zou H, McGarry TJ, Bernal T, Kirschner MW. Identification of a vertebrate sister-chromatid separation inhibitor involved in transformation and tumorigenesis. Science 1999;285:418-22
67. Csizmok V, Felli IC, Tompa P, et al. Structural and dynamic characterization of intrinsically disordered human securin by NMR spectroscopy. J Am Chem Soc 2008;130:16873-9
68. Hornig NC, Knowles PP, McDonald NQ, Uhlmann F. The dual mechanism of separase regulation by securin. Curr Biol 2002;12:973-82
69. Jallepalli PV, Waizenegger IC, Bunz F, et al. Securin is required for chromosomal stability in human cells. Cell 2001;105: 445-57
70. Barberis M. Sic1 as a timer of Clb cyclin waves in the yeast cell cycle-design principle of not just an inhibitor. FEBS J 2012;279: 3386-410
71. Borg M, Mittag T, Pawson T, et al. Polyelectrostatic interactions of disordered ligands suggest a physical basis for ultrasensitivity. Proc Natl Acad Sci U S A 2007;104:9650-5 .. On the basis of studies of the yeast cyclin-dependent kinase inhibitor Sic1 binding to Cdc4, the authors built a model that predicts a threshold number of phosphorylation sites for receptor- ligand binding, suggesting that ultrasensitivity in the Sic1-Cdc4 system may be driven at least, in part, by cumulative electrostatic interactions. The authors suggest that polyelectrostatic interactions may provide a simple yet powerful framework for understanding the modulation of protein interactions by multiple phosphorylation sites in disordered protein regions.
72. Pelka P, Ablack JN, Fonseca GJ, et al. Intrinsic structural disorder in adenovirus E1A: a viral molecular hub linking multiple diverse processes. J Virol 2008;82:7252-63
73. Chemes LB, Sanchez IE, Smal C, de Prat-Gay G. Targeting mechanism of the retinoblastoma tumor suppressor by a prototypical viral oncoprotein. Structural modularity, intrinsic disorder and phosphorylation of human papillomavirus E7. FEBS J 2010;277:973-88
74. Dosztanyi Z, Csizmok V, Tompa P, Simon I. IUPred: web server for the prediction of intrinsically unstructured regions of proteins based on estimated energy content. Bioinformatics 2005;21: 3433-4
75. Dosztanyi Z, Sandor M, Tompa P, Simon I. Prediction of protein disorder at the domain level. Curr Protein Pept Sci 2007;8:161-71
76. Uversky VN, Oldfield CJ, Dunker AK. Intrinsically disordered proteins in human diseases: introducing the D2 concept. Annu Rev Biophys 2008;37:215-46
77. Elledge SJ, Winston J, Harper JW. A question of balance: the role of cyclin-kinase inhibitors in development and tumorigenesis. Trends Cell Biol 1996;6: 388-92
78. Hall M, Peters G. Genetic alterations of cyclins, cyclin-dependent kinases, and Cdk inhibitors in human cancer. Adv Cancer Res 1996;68:67-108
79. Tsai T, Davalath S, Rankin C, et al. Tumor suppressor gene alteration in adult acute lymphoblastic leukemia (ALL). Analysis of retinoblastoma (Rb) and p53 gene expression in lymphoblasts of patients with de novo, relapsed, or refractory ALL treated in Southwest Oncology Group studies. Leukemia 1996;10:1901-10
80. Greenblatt MS, Bennett WP, Hollstein M, Harris CC. Mutations in the p53 tumor suppressor gene: clues to cancer etiology and molecular pathogenesis. Cancer Res 1994;54:4855-78
81. Crook T, Vousden KH. Interaction of HPV E6 with p53 and associated proteins. Biochem Soc Trans 1994;22:52-5
82. Ball AR Jr, Chen YY, Yokomori K. Mechanisms of cohesin-mediated gene regulation and lessons learned from cohesinopathies. Biochim Biophys Acta 2014;1839:191-202
83. Lapenna S, Giordano A. Cell cycle kinases as therapeutic targets for cancer. Nat Rev Drug Discov 2009;8:547-66
84. Vassilev LT, Vu BT, Graves B, et al. In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science 2004;303:844-8 .. The authors report the identification of potent and selective small-molecule antagonists of MDM2 and the confirmation of their mode of action through the crystal structures of complexes. These compounds bind MDM2 in the p53-binding pocket and activate the p53 pathway in cancer cells, leading to cell cycle arrest, apoptosis, and growth inhibition of human tumor xenografts in nude mice.
85. Sebolt-Leopold JS, English JM. Mechanisms of drug inhibition of signalling molecules. Nature 2006;441:457-62
86. Lagerstrom MC, Schioth HB. Structural diversity of G protein-coupled receptors and significance for drug discovery. Nat Rev Drug Discov 2008;7:339-57
87. Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 2001;46:3-26
88. Drews J. Drug discovery: a historical perspective. Science 2000;287:1960-4
89. Arkin M. Protein-protein interactions and cancer: small molecules going in for the kill. Curr Opin Chem Biol 2005;9:317-24
90. Arkin MR, Wells JA. Small-molecule inhibitors of protein-protein interactions: progressing towards the dream. Nat Rev Drug Discov 2004;3:301-17
91. Fuxreiter M, Simon I, Friedrich P, Tompa P. Preformed structural elements feature in partner recognition by intrinsically unstructured proteins. J Mol Biol 2004;338:1015-26
92. Oldfield CJ, Cheng Y, Cortese MS, et al. Coupled folding and binding with alpha-helix-forming molecular recognition elements. Biochemistry 2005;44:12454-70
93. Diella F, Haslam N, Chica C, et al. Understanding eukaryotic linear motifs and their role in cell signaling and regulation. Front Biosci 2008;13:6580-603
94. Tompa P, Davey NE, Gibson TJ, Babu MM. A million peptide motifs for the molecular biologist. Mol Cell 2014;55: 161-9
95. Cheng Y, LeGall T, Oldfield CJ, et al. Rational drug design via intrinsically disordered protein. Trends Biotechnol 2006;24:435-42 . In this article, the authors point out the importance of coupled binding and folding for protein-protein signaling interactions generally, and from this and associated observations, they develop a new strategy for identifying protein- protein interactions that would be particularly promising targets for modulation by small molecules.
96. Lane D, Levine A. p53 Research: the past thirty years and the next thirty years. Cold Spring Harb Perspect Biol 2010;2:a000893
97. Moll UM, Petrenko O. The MDM2-p53 interaction. Mol Cancer Res 2003;1:1001-8
98. Hammoudeh DI, Follis AV, Prochownik EV, Metallo SJ. Multiple independent binding sites for small-molecule inhibitors on the oncoprotein c-Myc. J Am Chem Soc 2009;131:7390-401
99. Metallo SJ. Intrinsically disordered proteins are potential drug targets. Curr Opin Chem Biol 2010;14:481-8 .. The binding of small molecules to IDPs is addressed in the article. The tendency of IDPs to bind multiple, specific protein targets is recapitulated in their interaction with small molecules, since the conformational flexibility of the peptide recognition element allows the plasticity to bind molecules with divergent structures with similar affinity. A strategy to develop small molecule binders of IDPs is introduced.
100. Hegyi H, Buday L, Tompa P. Intrinsic structural disorder confers cellular viability on oncogenic fusion proteins. PLoS Comput Biol 2009;5:e1000552
101. Erkizan HV, Kong Y, Merchant M, et al. A small molecule blocking oncogenic protein EWS-FLI1 interaction with RNA helicase A inhibits growth of Ewing's sarcoma. Nat Med 2009;15:750-6
102. Krishnan N, Koveal D, Miller DH, et al. Targeting the disordered C terminus of PTP1B with an allosteric inhibitor. Nat Chem Biol 2014;10:558-66
103. Wenthur CJ, Gentry PR, Mathews TP, Lindsley CW. Drugs for allosteric sites on receptors. Annu Rev Pharmacol Toxicol 2014;54:165-84
104. Tompa P. Multisteric regulation by structural disorder in modular signaling proteins: an extension of the concept of allostery. Chem Rev 2014;114:6715-32
105. Xue L, Wang P, Cao P, et al. Identification of extracellular signal-regulated kinase 1 (ERK1) direct substrates using stable isotope labeled kinase assay-linked phosphoproteomics. Mol Cell Proteomics 2014;13:3199-210
106. Xue L, Wang WH, Iliuk A, et al. Sensitive kinase assay linked with phosphoproteomics for identifying direct kinase substrates. Proc Natl Acad Sci USA 2012;109:5615-20
107. Li Y, Cross FR, Chait BT. Method for identifying phosphorylated substrates of specific cyclin/cyclin-dependent kinase complexes. Proc Natl Acad Sci USA 2014;111:11323-8
108. Blethrow JD, Glavy JS, Morgan DO, Shokat KM. Covalent capture of kinase-specific phosphopeptides reveals Cdk1-cyclin B substrates. Proc Natl Acad Sci USA 2008;105:1442-7
109. Archambault V, Chang EJ, Drapkin BJ, et al. Targeted proteomic study of the cyclin-Cdk module. Mol Cell 2004;14: 699-711
110. Pagliuca FW, Collins MO, Lichawska A, et al. Quantitative proteomics reveals the basis for the biochemical specificity of the cell-cycle machinery. Mol Cell 2011;43: 406-17 .. The authors performed a time-resolved analysis of the in vivo interactors of cyclins E1, A2 and B1. This revealed the temporal dynamics of cyclin function in which networks of cyclin-CDK interactions vary according to the type of cyclin and cell cycle stage. The results explain the temporal specificity of the cell cycle machinery, thereby providing a biochemical mechanism for the genetic requirement for multiple cyclins in vivo, and reveal how the actions of specific cyclins are coordinated to control the cell cycle.
111. Kettenbach AN, Schweppe DK, Faherty BK, et al. Quantitative phosphoproteomics identifies substrates and functional modules of Aurora and Polo-like kinase activities in mitotic cells. Sci Signal 2011;4:rs5
112. Smolka MB, Albuquerque CP, Chen SH, Zhou H. Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases. Proc Natl Acad Sci USA 2007;104: 10364-9
113. Matsuoka S, Ballif BA, Smogorzewska A, et al. ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage. Science 2007;316:1160-6
114. Selenko P, Wagner G. Looking into live cells with in-cell NMR spectroscopy. J Struct Biol 2007;158:244-53
115. Borcherds W, Theillet FX, Katzer A, et al. Disorder and residual helicity alter p53-Mdm2 binding affinity and signaling in cells. Nat Chem Biol 2014;10:1000-2