Melanoma: Targeting signaling pathways and RaLP

Expert Opinion on Therapeutic Targets

Volumn 13, Issue 1, 2009, Pages 93-104

  Pasini, Luigi ^a   Turco, Margherita Y ^a   Luzi, Lucilla ^a   Aladowicz, Ewa ^a   Fagiani, Ernesta ^b   Lanfrancone, Luisa ^a  

^a EUROPEAN INSTITUTE OF ONCOLOGY   (Italy)

^b Department of Clinical and Biological Research ^*  (Switzerland)

Author keywords

Metastatic melanoma; RaLP; Shc family; Targeted inhibitors

Indexed keywords

2 MORPHOLINO 8 PHENYLCHROMONE; 5 (4 BROMO 2 CHLOROANILINO) 4 FLUORO 1 METHYL 1H BENZIMIDAZOLE 6 CARBOHYDROXAMIC ACID 2 HYDROXYETHYL ESTER; ANTINEOPLASTIC AGENT; BEVACIZUMAB; CHIR 265; CILENGITIDE; CNTO 95; DACARBAZINE; IMATINIB; INTEGRIN; ISIS 2503; LONAFARNIB; MITOGEN ACTIVATED PROTEIN KINASE 1; MITOGEN ACTIVATED PROTEIN KINASE 3; MONOCLONAL ANTIBODY LM 609; N (2,3 DIHYDROXYPROPOXY) 3,4 DIFLUORO 2 (2 FLUORO 4 IODOANILINO)BENZAMIDE; PERIFOSINE; PHOSPHATIDYLINOSITOL 3 KINASE; PLX 4032; PROTEIN KINASE B; PROTEIN SHC; PROTEIN TYROSINE PHOSPHATASE SHP; PX 866; RAI LIKE PROTEIN; RAPAMYCIN; RX 0201; SORAFENIB; TEMSIROLIMUS; TIPIFARNIB; UNCLASSIFIED DRUG; VOLOCIXIMAB; WORTMANNIN; SHC4 PROTEIN, HUMAN;

ADJUVANT THERAPY; ANGIOGENESIS; APOPTOSIS; CANCER COMBINATION CHEMOTHERAPY; CANCER GROWTH; CELL PROLIFERATION; CELL SURVIVAL; CONSTIPATION; DRUG BIOAVAILABILITY; EPIDERMIS; HUMAN; LIFESPAN; MELANOMA; METASTASIS; NAUSEA; NONHUMAN; PROTEIN EXPRESSION; PROTEIN FUNCTION; PROTEIN TARGETING; PROTEIN TRANSPORT; REVIEW; SIGNAL TRANSDUCTION; SRC HOMOLOGY DOMAIN; TUMOR CELL; AMINO ACID SEQUENCE; ARTICLE; CHEMISTRY; DISEASE COURSE; METABOLISM; MOLECULAR GENETICS; PATHOLOGY; SEQUENCE HOMOLOGY;

AMINO ACID SEQUENCE; DISEASE PROGRESSION; HUMANS; MELANOMA; MOLECULAR SEQUENCE DATA; NEOPLASM METASTASIS; SEQUENCE HOMOLOGY, AMINO ACID; SHC SIGNALING ADAPTOR PROTEINS; SIGNAL TRANSDUCTION;

EID: 61449562203     PISSN: 14728222     EISSN: None     Source Type: Journal    
DOI: 10.1517/14728220802607363     Document Type: Review

References (101)

1. Liu V, Mihm MC. Pathology of malignant melanoma. Surg Clin North Am 2003 ; 83 (1): 31-60 (Pubitemid 36384359)
2. Miller AJ, Mihm MC. Melanoma. N Engl J Med 2006 ; 355 (1): 51-65
3. Scolyer RA, Murali R, McCarthy SW, Thompson JF. Pathologic examination of sentinel lymph nodes from melanoma patients. Semin Diagn Pathol 2008 ; 25 (2): 100-111 (Pubitemid 351885456)
4. Chudnovsky Y, Khavari PA, Adams AE. Melanoma genetics and the development of rational therapeutics. J Clin Invest 2005 ; 115 (4): 813-824 (Pubitemid 40489381)
5. Gogas HJ, Kirkwood JM, Sondak V. Chemotherapy for metastatic melanoma: time for a change? Cancer 2007 ; 109 (3): 455-464
6. Tawbi HA, Kirkwood JM. Management of metastatic melanoma. Semin Oncol 2007 ; 34 (6): 532-545 (Pubitemid 350236308)
7. Jemal A, Siegel R, Ward E, et al. Cancer statistics, 2008. CA Cancer J Clin 2008 ; 58 (2): 71-96
8. Soengas MS, Lowe SW. Apoptosis and melanoma chemoresistance. Oncogene 2003 ; 22 (20): 3138-3151 (Pubitemid 36713766)
9. Bedikian AY, Millward M, Pehamberger H, et al. Bcl-2 antisense (oblimersen sodium) plus dacarbazine in patients with advanced melanoma: the Oblimersen Melanoma Study Group. J Clin Oncol 2006; 24 (29): 4738-4745 (Pubitemid 46630937)
10. Grover R, Wilson GD. Bcl-2 expression in malignant melanoma and its prognostic signifi cance. Eur J Surg Oncol 1996; 22 (4): 347-349 (Pubitemid 26312873)
11. Pilla L, Valenti R, Marrari A, et al. Vaccination: role in metastatic melanoma. Expert Rev Anticancer Ther 2006 ; 6 (8): 1305-1318 (Pubitemid 44304698)
12. Tarhini AA, Agarwala SS. Interleukin-2 for the treatment of melanoma. Curr opin investig drugs 2005 ; 6 (12): 1234-1239 (Pubitemid 43009203)
13. Tarhini AA, Agarwala SS. Cutaneous melanoma: available therapy for metastatic disease. Dermatol Ther 2006 ; 19 (1): 19-25
14. Bennett DC. How to make a melanoma: what do we know of the primary clonal events? Pigment cell melanoma res 2008 ; 21 (1): 27-38
15. Hayward NK. Genetics of melanoma predisposition. Oncogene 2003 ; 22 (20): 3053-3062 (Pubitemid 36713755)
16. Crowson AN, Magro C, Miller AJ, Mihm MC. The molecular basis of melanomagenesis and the metastatic phenotype. Semin Oncol 2007; 34 (6): 476-490 (Pubitemid 350245678)
17. Giacinti C, Giordano A. RB and cell cycle progression. Oncogene 2006 ; 25 (38): 5220-5227 (Pubitemid 44299080)
18. Fargnoli MC, Argenziano G, Zalaudek I, Peris K. High- and low-penetrance cutaneous melanoma susceptibility genes. Expert Rev Anticancer Ther 2006; 6 (5): 657-670 (Pubitemid 44192883)
19. Sekulic A, Haluska P Jr, Miller AJ, et al. Malignant melanoma in the 21st century: the emerging molecular landscape. Mayo Clin Proc 2008 ; 83 (7): 825-846 (Pubitemid 351948695)
20. Cully M, You H, Levine AJ, Mak TW. Beyond PTEN mutations: the PI3K pathway as an integrator of multiple inputs during tumorigenesis. Nat Rev Cancer 2006; 6 (3): 184-192 (Pubitemid 43292562)
21. Mc Kay MM, Morrison DK. Integrating signals from RTKs to ERK/MAPK. Oncogene 2007 ; 26 (22): 3113-3121 (Pubitemid 46763008)
22. Polsky D, Cordon-Cardo C. Oncogenes in melanoma. Oncogene 2003 ; 22 (20): 3087-3091 (Pubitemid 36713760)
23. Haluska F, Pemberton T, Ibrahim N, Kalinsky K. The RTK/RAS/BRAF/PI3K pathways in melanoma: biology, small molecule inhibitors, and potential applications. Semin Oncol 2007; 34 (6): 546-554 (Pubitemid 350236312)
24. Guo W, Giancotti FG. Integrin signalling during tumour progression. Nat Rev Mol Cell Biol 2004; 5 (10): 816-826 (Pubitemid 39336275)
25. Fecher LA, Cummings SD, Keefe MJ, Alani RM. Toward a molecular classifi cation of melanoma. J Clin Oncol 2007; 25 (12): 1606-1620 (Pubitemid 46733089)
26. Gray-Schopfer V, Wellbrock C, Marais R. Melanoma biology and new targeted therapy. Nature 2007 ; 445 (7130): 851-857 (Pubitemid 46309849)
27. Fagiani E, Giardina G, Luzi L, et al. RaLP, a new member of the Src homology and collagen family, regulates cell migration and tumor growth of metastatic melanomas. Cancer Res 2007 ; 67 (7): 3064-3073 (Pubitemid 46724843)
28. Yoon S, Seger R. The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions. Growth Factors 2006 ; 24 (1): 21-44 (Pubitemid 43108774)
29. Huang C, Jacobson K, Schaller MD. MAP kinases and cell migration. J Cell Sci 2004 ; 117 (Pt 20): 4619-4628 (Pubitemid 39433429)
30. Fecher LA, Amaravadi RK, Flaherty KT. The MAPK pathway in melanoma. Curr opin Oncol 2008 ; 20 (2): 183-189 (Pubitemid 351317406)
31. Dhillon AS, Hagan S, Rath O, Kolch W. MAP kinase signalling pathways in cancer. Oncogene 2007 ; 26 (22): 3279-3290 (Pubitemid 46763021)
32. Hubbard SR, Miller WT. Receptor tyrosine kinases: mechanisms of activation and signaling. Curr Opin Cell Biol 2007 ; 19 (2): 117-123 (Pubitemid 46386407)
33. Kohno M, Pouyssegur J. Targeting the ERK signaling pathway in cancer therapy. Ann Med 2006 ; 38 (3): 200-211 (Pubitemid 43675585)
34. Wan PT, Garnett MJ, Roe SM, et al. Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF. Cell 2004 ; 116 (6): 855-867 (Pubitemid 38410730)
35. Gray-Schopfer V, Da Rocha Dias S, Marais R. The role of B-RAF in melanoma. Cancer Metastasis Rev 2005 ; 24 (1): 165-183 (Pubitemid 40576070)
36. Wellbrock C, Ogilvie L, Hedley D, et al. V599EB-RAF is an oncogene in melanocytes. Cancer Res 2004 ; 64 (7): 2338-2342 (Pubitemid 38523884)
37. Cohen C, Zavala-Pompa A, Sequeira JH, et al. Mitogen-actived protein kinase activation is an early event in melanoma progression. Clin Cancer Res 2002 ; 8 (12): 3728-3733 (Pubitemid 35424762)
38. Grichnik JM. Kit and melanocyte migration. J Invest Dermatol 2006 ; 126 (5): 945-947
39. Roskoski R. Signaling by Kit protein-tyrosine kinase-the stem cell factor receptor. Biochem Biophys Res Commun 2005 ; 337 (1): 1-13 (Pubitemid 41393751)
40. Satyamoorthy K, Li G, Gerrero MR, et al. Constitutive mitogen-activated protein kinase activation in melanoma is mediated by both BRAF mutations and autocrine growth factor stimulation. Cancer Res 2003 ; 63 (4): 756-759 (Pubitemid 36231971)
41. Paez J, Sellers WR. PI3K/PTEN/AKT pathway. A critical mediator of oncogenic signaling. Cancer Treat Res 2003; 115 : 145-167
42. Li G, Satyamoorthy K, Herlyn M. N-cadherin-mediated intercellular interactions promote survival and migration of melanoma cells. Cancer Res 2001; 61 (9): 3819-3825 (Pubitemid 32694999)
43. Kang SS, Kwon T, Kwon DY, Do SI. Akt protein kinase enhances human telomerase activity through phosphorylation of telomerase reverse transcriptase subunit. J Biol Chem 1999 ; 274 (19): 13085-13090 (Pubitemid 129518508)
44. Li G, Kalabis J, Xu X, et al. Reciprocal regulation of MelCAM and AKT in human melanoma. Oncogene 2003 ; 22 (44): 6891-6899 (Pubitemid 37386689)
45. Govindarajan B, Sligh JE, Vincent BJ, et al. Overexpression of Akt converts radial growth melanoma to vertical growth melanoma. J Clin Invest 2007; 117 (3): 719-729 (Pubitemid 46348530)
46. Cheung M, Sharma A, Madhunapantula SV, Robertson GP. Akt3 and mutant V600EB-Raf cooperate to promote early melanoma development. Canaer Res 2008 ; 68 (9): 3429-3439
47. Davies MA, Stemke-Hale K, Tellez C, et al. A novel AKT3 mutation in melanoma tumours and cell lines. Br J Cancer 2008 ; 99 (8): 1265-1268
48. Robertson GP. Functional and therapeutic signifi cance of Akt deregulation in malignant melanoma. Cancer Metastasis Rev 2005 ; 24 (2): 273-285 (Pubitemid 40922799)
49. Wu H, Goel V, Haluska F. PTEN signaling pathways in melanoma. Oncogene 2003 ; 22 (20): 3113-3122 (Pubitemid 36713763)
50. Gu J, Tamura M, Pankov R, et al. Shc and FAK differentially regulate cell motility and directionality modulated by PTEN. J Cell Biol 1999 ; 146 (2): 389-403 (Pubitemid 29369775)
51. Stahl JM, Cheung M, Sharma A, et al. Loss of PTEN promotes tumor development in malignant melanoma. Cancer Res 2003 ; 63 (11): 2881-2890 (Pubitemid 36667160)
52. Van Der Flier A, Sonnenberg A. Function and interactions of integrins. Cell Tissue Res 2001 ; 305 (3): 285-298 (Pubitemid 34212122)
53. Hehlgans S, Haase M, Cordes N. Signalling via integrins: implications for cell survival and anticancer strategies. Biochim Biophys Acta 2007;1775(1):163-180 (Pubitemid 44881199)
54. Schwartz MA, Horwitz AR. Integrating adhesion, protrusion, and contraction during cell migration. Cell 2006 ; 125 (7): 1223-1225 (Pubitemid 43929100)
55. Giancotti FG, Ruoslahti E. Integrin signaling. Science 1999 ; 285 (5430): 1028-32
56. Eliceiri BP, Cheresh DA. Adhesion events in angiogenesis. Curr Opin Cell Biol 2001; 13 (5): 563-568
57. Howe AK, Aplin AE, Juliano RL. Anchorage-dependent ERK signaling-mechanisms and consequences. Curr Opin Genet Dev 2002 ; 12 (1): 30-35
58. Kuphal S, Bauer R, Bosserhoff AK. Integrin signaling in malignant melanoma. Cancer Metastasis Rev 2005 ; 24 (2): 195-222 (Pubitemid 40922795)
59. Barberis L, Wary KK, Fiucci G, et al. Distinct roles of the adaptor protein Shc and focal adhesion kinase in integrin signaling to ERK. J Biol Chem 2000; 275 (47): 36532-36540 (Pubitemid 32002050)
60. Luzi L, Confalonieri S, Di Fiore PP, Pelicci PG. Evolution of Shc functions from nematode to human. Curr Opin Genet Dev 2000 ; 10 (6): 668-674
61. Jones N, Hardy WR, Friese MB, et al. Analysis of a Shc family adaptor protein, ShcD/Shc4, that associates with muscle-specifi c kinase. Mol Cell Biol 2007 ; 27 (13): 4759-4773 (Pubitemid 47016129)
62. Migliaccio E, Mele S, Salcini AE, et al. Opposite effects of the p52shc/p46shc and p66shc splicing isoforms on the EGF receptor-MAP kinase-fos signalling pathway. EMBO J 1997 ; 16 (4): 706-716 (Pubitemid 27089518)
63. Pelicci G, Troglio F, Bodini A, et al. The neuron-specifi c Rai (ShcC) adaptor protein inhibits apoptosis by coupling Ret to the phosphatidylinositol 3-kinase/Akt signaling pathway. Mol Cell Biol 2002;22(20):7351-7363
64. Kojima T, Yoshikawa Y, Takada S, et al. Genomic organization of the Shc-related phosphotyrosine adapters and characterization of the full-length Sck/ShcB: specifi c association of p68-Sck/ShcB with pp135. Biochem Biophys Res Commun 2001; 284 (4): 1039-1047 (Pubitemid 32924753)
65. Cattaneo E, Pelicci PG. Emerging roles for SH2/PTB-containing Shc adaptor proteins in the developing mammalian brain. Trends Neurosci 1998 ; 21 (11): 476-481 (Pubitemid 28506138)
66. Nakamura T, Muraoka S, Sanokawa R, Mori N. N-Shc and Sck, two neuronally expressed Shc adapter homologs. Their differential regional expression in the brain and roles in neurotrophin and Src signaling. J Biol Chem 1998 ; 273 (12): 6960-6967 (Pubitemid 28160362)
67. Sakai R, Henderson JT, O'Bryan JP, et al. The mammalian ShcB and ShcC phosphotyrosine docking proteins function in the maturation of sensory and sympathetic neurons. Neuron 2000; 28 (3): 819-833 (Pubitemid 32038074)
68. Lai KM, Pawson T. The ShcA phosphotyrosine docking protein sensitizes cardiovascular signaling in the mouse embryo. Genes Dev 2000; 14 (9): 1132-1145 (Pubitemid 30324426)
69. Migliaccio E, Giorgio M, Mele S, et al. The p66shc adaptor protein controls oxidative stress response and life span in mammals. Nature 1999 ; 402 (6759): 309-313 (Pubitemid 129516269)
70. Ravichandran KS. Signaling via Shc family adapter proteins. Oncogene 2001 ; 20 (44): 6322-6330 (Pubitemid 32977841)
71. Trinei M, Giorgio M, Cicalese A, et al. A p53-p66Shc signalling pathway controls intracellular redox status, levels of oxidation-damaged DNA and oxidative stress-induced apoptosis. Oncogene 2002 ; 21 (24): 3872-3878 (Pubitemid 34620855)
72. Pelicci G, Lanfrancone L, Grignani F, et al. A novel transforming protein (SHC) with an SH2 domain is implicated in mitogenic signal transduction. Cell 1992;70(1):93-104
73. Troglio F, Echart C, Gobbi A, et al. The Rai (Shc C) adaptor protein regulates the neuronal stress response and protects against cerebral ischemia. Proc Natl Acad Sci USA 2004 ; 101 (43): 15476-15481 (Pubitemid 39441566)
74. Ursini-Siegel J, Muller WJ. The ShcA adaptor protein is a critical regulator of breast cancer progression. Cell Cycle 2008 ; 7 (13): 1936-1943 (Pubitemid 352030337)
75. Grossman SR, Lyle S, Resnick MB, et al. p66 Shc tumor levels show a strong prognostic correlation with disease outcome in stage IIA colon cancer. Clin Cancer Res 2007 ; 13 (19): 5798-5804 (Pubitemid 47583903)
76. Terui E, Matsunaga T, Yoshida H, et al. Shc family expression in neuroblastoma: high expression of shcC is associated with a poor prognosis in advanced neuroblastoma. Clin Cancer Res 2005 ; 11 (9): 3280-3287 (Pubitemid 40627876)
77. Schlessinger J, Lemmon MA. SH2 and PTB domains in tyrosine kinase signaling. Sci STKE 2003; 2003 (191): RE12
78. Sebolt-Leopold J, English JM. Mechanisms of drug inhibition of signalling molecules. Nature 2006 ; 441 (7092): 457-462 (Pubitemid 44050141)
79. Kalinsky K, Haluska F. Novel inhibitors in the treatment of metastatic melanoma. Expert Rev Anticancer Ther 2007 ; 7 (5): 715-724 (Pubitemid 46805566)
80. Meier F, Busch S, Lasithiotakis KG, et al. Combined targeting of MAPK and AKT signalling pathways is a promising strategy for melanoma treatment. Br J Dermatol 2007 ; 156 (6): 1204-1213 (Pubitemid 47029801)
81. Smalley KS, Haass NK, Brafford PA, et al. Multiple signaling pathways must be targeted to overcome drug resistance in cell lines derived from melanoma metastases. Mol Cancer Ther 2006 ; 5 (5): 1136-1144 (Pubitemid 43881305)
82. Flaherty KT. Chemotherapy and targeted therapy combinations in advanced melanoma. Clin Cancer Res 2006 ; 12 (7 Pt 2): s2366-70
83. Gogas HJ, Kirkwood JM, Sondak VK. Chemotherapy for metastatic melanoma: time for a change? Cancer 2007 ; 109 (3): 455-464
84. Eisen T, Ahmad T, Flaherty KT, et al. Sorafenib in advanced melanoma: a Phase II randomised discontinuation trial analysis. Br J Cancer 2006 ; 95 (5): 581-586 (Pubitemid 44325897)
85. Wilhelm SM, Carter C, Tang L, et al. BAY 43-9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor progression and angiogenesis. Cancer Res 2004 ; 64 (19): 7099-7109 (Pubitemid 39331023)
86. Lorusso PM, Adjei AA, Varterasian M, et al. Phase I and pharmacodynamic study of the oral MEK inhibitor CI-1040 in patients with advanced malignancies. J Clin Oncol 2005 ; 23 (23): 5281-5293 (Pubitemid 46206982)
87. Wyman K, Atkins MB, Prieto V, et al. Multicenter Phase II trial of high-dose imatinib mesylate in metastatic melanoma: signifi cant toxicity with no clinical effi cacy. Cancer 2006 ; 106 (9): 2005-2011 (Pubitemid 43673217)
88. Jaissle GB, Ulmer A, Henke-Fahle S, et al. Suppression of melanoma-associated neoangiogenesis by bevacizumab. Arch Dermatol 2008 ; 144 (4): 525-527 (Pubitemid 351581110)
89. Bedogni B, Welford SM, Kwan AC, et al. Inhibition of phosphatidylinositol-3-kinase and mitogen-activated protein kinase kinase 1/2 prevents melanoma development and promotes melanoma regression in the transgenic TPRas mouse model. Mol Cancer Ther 2006 ; 5 (12): 3071-3077 (Pubitemid 46092048)
90. Hennessy BT, Smith D, Ram PT, et al. Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat rev Drug discov 2005 ; 4 (12): 988-1004
91. Ihle NT, Williams R, Chow S, et al. Molecular pharmacology and antitumor activity of PX-866, a novel inhibitor of phosphoinositide-3-kinase signaling. Mol Cancer Ther 2004 ; 3 (7): 763-772 (Pubitemid 39193758)
92. Ernst DS, Eisenhauer E, Waiaman N, et al. Phase II study of perifosine in previously untreated patients with metastatic melanoma. Invest new drugs 2005 ; 23 (6): 569-575 (Pubitemid 43162477)
93. Margolin K, Longmate J, Baratta T, et al. CCI-779 in metastatic melanoma: a Phase II trial of the California Cancer Consortium. Cancer 2005 ; 104 (5): 1045-1048 (Pubitemid 41170233)
94. Mousa SA. Cell adhesion molecules: potential therapeutic & diagnostic implications. Mol Biotechnol 2008 ; 38 (1): 33-40
95. Eskens FA, Dumez H, Hoekstra R, et al. Phase I and pharmacokinetic study of continuous twice weekly intravenous administration of Cilengitide (EMD 121974), a novel inhibitor of the integrins α v β 3 and α v β 5 in patients with advanced solid tumours. Eur J Cancer 2003 ; 39 (7): 917-926 (Pubitemid 36428623)
96. Posey JA, Khazaeli MB, Del Grosso A, et al. A pilot trial of Vitaxin, a humanized anti-vitronectin receptor (anti α v β 3) antibody in patients with metastatic cancer. Cancer Biother Radiopharm 2001 ; 16 (2): 125-132 (Pubitemid 34121636)
97. Hoek KS, Eichhoff OM, Schlegel NC, et al. In vivo switching of human melanoma cells between proliferative and invasive states. Cancer Res 2008 ; 68 (3): 650-656
98. Bittner M, Meltzer P, Chen Y, et al. Molecular classifi cation of cutaneous malignant melanoma by gene expression profi ling. Nature 2000 ; 406 (6795): 536-540 (Pubitemid 30625740)
99. Winnepenninckx V, Van Den Oord JJ. Gene expression profi ling and clinical outcome in melanoma: in search of novel prognostic factors. Expert Rev Anticancer Ther 2007 ; 7 (11): 1611-1631 (Pubitemid 350303016)
100. Molnár V, Tamási V, Bakos B, et al. Changes in miRNA expression in solid tumors: an miRNA profiling in melanomas. Semin Cancer Biol 2008 ; 18 (2): 111-122
101. Ursini-Siegel J, Hardy WR, Zuo D, et al. ShcA signalling is essential for tumour progression in mouse models of human breast cancer. EMBO J 2008 ; 27 (6): 910-920