Pathway choice in DNA double strand break repair: Observations of a balancing act

Genome Integrity

Volumn 3, Issue , 2012, Pages

  Brandsma, Inger ^a   Gent, Dik C ^a  

^a ERASMUS MEDICAL CENTER   (Netherlands)

Author keywords

DNA repair assays; DSB repair; HR; NHEJ; PARP inhibitors

Indexed keywords

BRCA1 PROTEIN; DOUBLE STRANDED DNA; ETOPOSIDE; OLAPARIB; PROTEIN P53;

CATALYSIS; CELL CYCLE G2 PHASE; CELL CYCLE S PHASE; CHROMOSOME TRANSLOCATION; CYTOTOXICITY; DNA END JOINING REPAIR; DNA REPAIR; DNA STRAND BREAKAGE; GENE INTERACTION; HETEROZYGOSITY; HOMOLOGOUS RECOMBINATION; IONIZING RADIATION; MOLECULAR DYNAMICS; NONHUMAN; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; PROTEIN PROTEIN INTERACTION; REVIEW; SEQUENCE ALIGNMENT; SEQUENCE HOMOLOGY; SIGNAL TRANSDUCTION; SUMOYLATION; TRANSPOSON; UBIQUITINATION; VDJ RECOMBINATION;

EID: 84873440196     PISSN: None     EISSN: 20419414     Source Type: Journal    
DOI: 10.1186/2041-9414-3-9     Document Type: Review

References (81)

1. Friedberg ECW, Siede GC, Wood RD, Schultz RA, Ellenberger T. DNA repair and mutagenesis 2006, Washington, USA: ASM press.
2. Ciccia A, Elledge SJ. The DNA damage response: making it safe to play with knives. Mol Cell 2010, 40:179-204. 10.1016/j.molcel.2010.09.019, 2988877, 20965415.
3. Lieber MR. The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway. Annu Rev Biochem 2010, 79:181-211. 10.1146/annurev.biochem.052308.093131, 3079308, 20192759.
4. Ahnesorg P, Smith P, Jackson SP. XLF interacts with the XRCC4-DNA ligase IV complex to promote DNA nonhomologous end-joining. Cell 2006, 124:301-313. 10.1016/j.cell.2005.12.031, 16439205.
5. Buck D, Malivert L, de Chasseval R, Barraud A, Fondaneche MC, Sanal O, Plebani A, Stephan JL, Hufnagel M, le Deist F, et al. Cernunnos, a novel nonhomologous end-joining factor, is mutated in human immunodeficiency with microcephaly. Cell 2006, 124:287-299. 10.1016/j.cell.2005.12.030, 16439204.
6. Heyer WD, Ehmsen KT, Liu J. Regulation of homologous recombination in eukaryotes. Annu Rev Genet 2010, 44:113-139. 10.1146/annurev-genet-051710-150955, 20690856.
7. Stracker TH, Petrini JH. The MRE11 complex: starting from the ends. Nat Rev Mol Cell Biol 2011, 12:90-103. 10.1038/nrm3047, 21252998.
8. Limbo O, Chahwan C, Yamada Y, de Bruin RA, Wittenberg C, Russell P. Ctp1 is a cell-cycle-regulated protein that functions with Mre11 complex to control double-strand break repair by homologous recombination. Mol Cell 2007, 28:134-146. 10.1016/j.molcel.2007.09.009, 2066204, 17936710.
9. Sartori AA, Lukas C, Coates J, Mistrik M, Fu S, Bartek J, Baer R, Lukas J, Jackson SP. Human CtIP promotes DNA end resection. Nature 2007, 450:509-514. 10.1038/nature06337, 2409435, 17965729.
10. Wyman C, Kanaar R. DNA double-strand break repair: all's well that ends well. Annu Rev Genet 2006, 40:363-383. 10.1146/annurev.genet.40.110405.090451, 16895466.
11. Sugiyama T, Zaitseva EM, Kowalczykowski SC. A single-stranded DNA-binding protein is needed for efficient presynaptic complex formation by the Saccharomyces cerevisiae Rad51 protein. J Biol Chem 1997, 272:7940-7945. 10.1074/jbc.272.12.7940, 9065463.
12. Pardo B, Gomez-Gonzalez B, Aguilera A. DNA repair in mammalian cells: DNA double-strand break repair: how to fix a broken relationship. Cell Mol Life Sci 2009, 66:1039-1056. 10.1007/s00018-009-8740-3, 19153654.
13. Maher RL, Branagan AM, Morrical SW. Coordination of DNA replication and recombination activities in the maintenance of genome stability. J Cell Biochem 2011, 112:2672-2682. 10.1002/jcb.23211, 3178728, 21647941.
14. Polo SE, Jackson SP. Dynamics of DNA damage response proteins at DNA breaks: a focus on protein modifications. Genes Dev 2011, 25:409-433. 10.1101/gad.2021311, 3049283, 21363960.
15. Kim JS, Krasieva TB, Kurumizaka H, Chen DJ, Taylor AM, Yokomori K. Independent and sequential recruitment of NHEJ and HR factors to DNA damage sites in mammalian cells. J Cell Biol 2005, 170:341-347. 10.1083/jcb.200411083, 2171485, 16061690.
16. Mari PO, Florea BI, Persengiev SP, Verkaik NS, Bruggenwirth HT, Modesti M, Giglia-Mari G, Bezstarosti K, Demmers JA, Luider TM, et al. Dynamic assembly of end-joining complexes requires interaction between Ku70/80 and XRCC4. Proc Natl Acad Sci USA 2006, 103:18597-18602. 10.1073/pnas.0609061103, 1693708, 17124166.
17. Stark JM, Pierce AJ, Oh J, Pastink A, Jasin M. Genetic steps of mammalian homologous repair with distinct mutagenic consequences. Mol Cell Biol 2004, 24:9305-9316. 10.1128/MCB.24.21.9305-9316.2004, 522275, 15485900.
18. Audebert M, Salles B, Calsou P. Involvement of poly(ADP-ribose) polymerase-1 and XRCC1/DNA ligase III in an alternative route for DNA double-strand breaks rejoining. J Biol Chem 2004, 279:55117-55126. 10.1074/jbc.M404524200, 15498778.
19. Wang M, Wu W, Wu W, Rosidi B, Zhang L, Wang H, Iliakis G. PARP-1 and Ku compete for repair of DNA double strand breaks by distinct NHEJ pathways. Nucleic Acids Res 2006, 34:6170-6182. 10.1093/nar/gkl840, 1693894, 17088286.
20. Boboila C, Oksenych V, Gostissa M, Wang JH, Zha S, Zhang Y, Chai H, Lee CS, Jankovic M, Saez LM, et al. Robust chromosomal DNA repair via alternative end-joining in the absence of X-ray repair cross-complementing protein 1 (XRCC1). Proc Natl Acad Sci USA 2012, 109:2473-2478. 10.1073/pnas.1121470109, 3289296, 22308491.
21. Verkaik NS, Esveldt-van Lange RE, van Heemst D, Bruggenwirth HT, Hoeijmakers JH, Hoeijmakers MZ, van Gent DC. Different types of V(D)J recombination and end-joining defects in DNA double-strand break repair mutant mammalian cells. Eur J Immunol 2002, 32:701-709. 10.1002/1521-4141(200203)32:3<701::AID-IMMU701>3.0.CO;2-T, 11870614.
22. Mao Z, Bozzella M, Seluanov A, Gorbunova V. Comparison of nonhomologous end joining and homologous recombination in human cells. DNA Repair (Amst) 2008, 7:1765-1771.
23. Guirouilh-Barbat J, Huck S, Bertrand P, Pirzio L, Desmaze C, Sabatier L, Lopez BS. Impact of the KU80 pathway on NHEJ-induced genome rearrangements in mammalian cells. Mol Cell 2004, 14:611-623. 10.1016/j.molcel.2004.05.008, 15175156.
24. Coleman KA, Greenberg RA. The BRCA1-RAP80 complex regulates DNA repair mechanism utilization by restricting end resection. J Biol Chem 2011, 286:13669-13680. 10.1074/jbc.M110.213728, 3075711, 21335604.
25. Schulte-Uentrop L, El-Awady RA, Schliecker L, Willers H, Dahm-Daphi J. Distinct roles of XRCC4 and Ku80 in non-homologous end-joining of endonuclease- and ionizing radiation-induced DNA double-strand breaks. Nucleic Acids Res 2008, 36:2561-2569. 10.1093/nar/gkn094, 2377445, 18332040.
26. van Heemst D, Brugmans L, Verkaik NS, van Gent DC. End-joining of blunt DNA double-strand breaks in mammalian fibroblasts is precise and requires DNA-PK and XRCC4. DNA Repair (Amst) 2004, 3:43-50.
27. Yant SR, Kay MA. Nonhomologous-end-joining factors regulate DNA repair fidelity during Sleeping Beauty element transposition in mammalian cells. Mol Cell Biol 2003, 23:8505-8518. 10.1128/MCB.23.23.8505-8518.2003, 262663, 14612396.
28. Pan-Hammarstrom Q, Jones AM, Lahdesmaki A, Zhou W, Gatti RA, Hammarstrom L, Gennery AR, Ehrenstein MR. Impact of DNA ligase IV on nonhomologous end joining pathways during class switch recombination in human cells. J Exp Med 2005, 201:189-194. 10.1084/jem.20040772, 2212791, 15657289.
29. Van Der Burg M, Verkaik NS, den Dekker AT, Barendregt BH, Pico-Knijnenburg I, Tezcan I, vanDongen JJ, van Gent DC. Defective Artemis nuclease is characterized by coding joints with microhomology in long palindromic-nucleotide stretches. Eur J Immunol 2007, 37:3522-3528. 10.1002/eji.200737624, 18034425.
30. Lee GS, Neiditch MB, Salus SS, Roth DB. RAG proteins shepherd double-strand breaks to a specific pathway, suppressing error-prone repair, but RAG nicking initiates homologous recombination. Cell 2004, 117:171-184. 10.1016/S0092-8674(04)00301-0, 15084256.
31. Pierce AJ, Johnson RD, Thompson LH, Jasin M. XRCC3 promotes homology-directed repair of DNA damage in mammalian cells. Genes Dev 1999, 13:2633-2638. 10.1101/gad.13.20.2633, 317094, 10541549.
32. Hartlerode A, Odate S, Shim I, Brown J, Scully R. Cell cycle-dependent induction of homologous recombination by a tightly regulated I-SceI fusion protein. PLoS One 2011, 6:e16501. 10.1371/journal.pone.0016501, 3052302, 21408059.
33. Conrad S, Kunzel J, Lobrich M. Sister chromatid exchanges occur in G2-irradiated cells. Cell Cycle 2011, 10:222-228. 10.4161/cc.10.2.14639, 3048794, 21224723.
34. Lobrich M, Shibata A, Beucher A, Fisher A, Ensminger M, Goodarzi AA, Barton O, Jeggo PA. gammaH2AX foci analysis for monitoring DNA double-strand break repair: strengths, limitations and optimization. Cell Cycle 2010, 9:662-669. 10.4161/cc.9.4.10764, 20139725.
35. Karanam K, Kafri R, Loewer A, Lahav G. Quantitative live cell imaging reveals a gradual shift between DNA repair mechanisms and a maximal Use of HR in Mid S phase. Mol Cell 2012, 47:320-329. 10.1016/j.molcel.2012.05.052, 22841003.
36. Beucher A, Birraux J, Tchouandong L, Barton O, Shibata A, Conrad S, Goodarzi AA, Krempler A, Jeggo PA, Lobrich M. ATM and Artemis promote homologous recombination of radiation-induced DNA double-strand breaks in G2. EMBO J 2009, 28:3413-3427. 10.1038/emboj.2009.276, 2752027, 19779458.
37. Lamarche BJ, Orazio NI, Weitzman MD. The MRN complex in double-strand break repair and telomere maintenance. FEBS Lett 2010, 584:3682-3695. 10.1016/j.febslet.2010.07.029, 2946096, 20655309.
38. Huertas P, Jackson SP. Human CtIP mediates cell cycle control of DNA end resection and double strand break repair. J Biol Chem 2009, 284:9558-9565. 10.1074/jbc.M808906200, 2666608, 19202191.
39. Yun MH, Hiom K. CtIP-BRCA1 modulates the choice of DNA double-strand-break repair pathway throughout the cell cycle. Nature 2009, 459:460-463. 10.1038/nature07955, 2857324, 19357644.
40. Germani A, Prabel A, Mourah S, Podgorniak MP, Di Carlo A, Ehrlich R, Gisselbrecht S, Varin-Blank N, Calvo F, Bruzzoni-Giovanelli H. SIAH-1 interacts with CtIP and promotes its degradation by the proteasome pathway. Oncogene 2003, 22:8845-8851. 10.1038/sj.onc.1206994, 14654780.
41. Falck J, Forment JV, Coates J, Mistrik M, Lukas J, Bartek J, Jackson SP. CDK targeting of NBS1 promotes DNA-end resection, replication restart and homologous recombination. EMBO Rep 2012, 13:561-568. 10.1038/embor.2012.58, 22565321.
42. Cortes-Ledesma F, de Piccoli G, Haber JE, Aragon L, Aguilera A. SMC proteins, new players in the maintenance of genomic stability. Cell Cycle 2007, 6:914-918. 10.4161/cc.6.8.4107, 17404505.
43. Covo S, Westmoreland JW, Gordenin DA, Resnick MA. Cohesin Is limiting for the suppression of DNA damage-induced recombination between homologous chromosomes. PLoS Genet 2010, 6:e1001006. 10.1371/journal.pgen.1001006, 2895640, 20617204.
44. Spitzner JR, Chung IK, Gootz TD, McGuirk PR, Muller MT. Analysis of eukaryotic topoisomerase II cleavage sites in the presence of the quinolone CP-115,953 reveals drug-dependent and -independent recognition elements. Mol Pharmacol 1995, 48:238-249.
45. Shibata A, Conrad S, Birraux J, Geuting V, Barton O, Ismail A, Kakarougkas A, Meek K, Taucher-Scholz G, Lobrich M, Jeggo PA. Factors determining DNA double-strand break repair pathway choice in G2 phase. EMBO J 2011, 30:1079-1092. 10.1038/emboj.2011.27, 3061033, 21317870.
46. Hada M, Georgakilas AG. Formation of clustered DNA damage after high-LET irradiation: a review. J Radiat Res 2008, 49:203-210. 10.1269/jrr.07123, 18413977.
47. Goodarzi AA, Noon AT, Deckbar D, Ziv Y, Shiloh Y, Lobrich M, Jeggo PA. ATM signaling facilitates repair of DNA double-strand breaks associated with heterochromatin. Mol Cell 2008, 31:167-177. 10.1016/j.molcel.2008.05.017, 18657500.
48. Ziv Y, Bielopolski D, Galanty Y, Lukas C, Taya Y, Schultz DC, Lukas J, Bekker-Jensen S, Bartek J, Shiloh Y. Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway. Nat Cell Biol 2006, 8:870-876. 10.1038/ncb1446, 16862143.
49. Goodarzi AA, Kurka T, Jeggo PA. KAP-1 phosphorylation regulates CHD3 nucleosome remodeling during the DNA double-strand break response. Nat Struct Mol Biol 2011, 18:831-839. 10.1038/nsmb.2077, 21642969.
50. Costelloe T, Louge R, Tomimatsu N, Mukherjee B, Martini E, Khadaroo B, Dubois K, Wiegant WW, Thierry A, Burma S, et al. The yeast Fun30 and human SMARCAD1 chromatin remodellers promote DNA end resection. Nature 2012, 489(7417):581-584. 10.1038/nature11353, 22960744.
51. Sonoda E, Hochegger H, Saberi A, Taniguchi Y, Takeda S. Differential usage of non-homologous end-joining and homologous recombination in double strand break repair. DNA Repair (Amst) 2006, 5:1021-1029.
52. Rappold I, Iwabuchi K, Date T, Chen J. Tumor suppressor p53 binding protein 1 (53BP1) is involved in DNA damage-signaling pathways. J Cell Biol 2001, 153:613-620. 10.1083/jcb.153.3.613, 2190566, 11331310.
53. Noon AT, Shibata A, Rief N, Lobrich M, Stewart GS, Jeggo PA, Goodarzi AA. 53BP1-dependent robust localized KAP-1 phosphorylation is essential for heterochromatic DNA double-strand break repair. Nat Cell Biol 2010, 12:177-184. 10.1038/ncb2017, 20081839.
54. Bothmer A, Robbiani DF, Feldhahn N, Gazumyan A, Nussenzweig A, Nussenzweig MC. 53BP1 regulates DNA resection and the choice between classical and alternative end joining during class switch recombination. J Exp Med 2010, 207:855-865. 10.1084/jem.20100244, 2856023, 20368578.
55. Difilippantonio S, Gapud E, Wong N, Huang CY, Mahowald G, Chen HT, Kruhlak MJ, Callen E, Livak F, Nussenzweig MC, et al. 53BP1 facilitates long-range DNA end-joining during V(D)J recombination. Nature 2008, 456:529-533. 10.1038/nature07476, 18931658.
56. Dimitrova N, Chen YC, Spector DL, de Lange T. 53BP1 promotes non-homologous end joining of telomeres by increasing chromatin mobility. Nature 2008, 456:524-528. 10.1038/nature07433, 2613650, 18931659.
57. Moynahan ME, Chiu JW, Koller BH, Jasin M. Brca1 controls homology-directed DNA repair. Mol Cell 1999, 4:511-518. 10.1016/S1097-2765(00)80202-6, 10549283.
58. Chen JJ, Silver D, Cantor S, Livingston DM, Scully R. BRCA1, BRCA2, and Rad51 operate in a common DNA damage response pathway. Cancer Res 1999, 59:1752s-1756s.
59. Ohta T, Sato K, Wu W. The BRCA1 ubiquitin ligase and homologous recombination repair. FEBS Lett 2011, 585:2836-2844. 10.1016/j.febslet.2011.05.005, 21570976.
60. Drost R, Bouwman P, Rottenberg S, Boon U, Schut E, Klarenbeek S, Klijn C, Van Der Heijden I, Van Der Gulden H, Wientjens E, et al. BRCA1 RING function is essential for tumor suppression but dispensable for therapy resistance. Cancer Cell 2011, 20:797-809. 10.1016/j.ccr.2011.11.014, 22172724.
61. Chen L, Nievera CJ, Lee AY, Wu AY. Cell cycle-dependent complex formation of BRCA1.CtIP.MRN is important for DNA double-strand break repair. J Biol Chem 2008, 283:7713-7720. 10.1074/jbc.M710245200, 18171670.
62. Kim H, Chen J, Yu X. Ubiquitin-binding protein RAP80 mediates BRCA1-dependent DNA damage response. Science 2007, 316:1202-1205. 10.1126/science.1139621, 17525342.
63. Sobhian B, Shao G, Lilli DR, Culhane AC, Moreau LA, Xia B, Livingston DM, Greenberg RA. RAP80 targets BRCA1 to specific ubiquitin structures at DNA damage sites. Science 2007, 316:1198-1202. 10.1126/science.1139516, 2706583, 17525341.
64. Wang B, Matsuoka S, Ballif BA, Zhang D, Smogorzewska A, Gygi SP, Elledge SJ. Abraxas and RAP80 form a BRCA1 protein complex required for the DNA damage response. Science 2007, 316:1194-1198. 10.1126/science.1139476, 17525340.
65. Cao L, Xu X, Bunting SF, Liu J, Wang RH, Cao LL, Wu JJ, Peng TN, Chen J, Nussenzweig A, et al. A selective requirement for 53BP1 in the biological response to genomic instability induced by Brca1 deficiency. Mol Cell 2009, 35:534-541. 10.1016/j.molcel.2009.06.037, 3392030, 19716796.
66. Bouwman P, Aly A, Escandell JM, Pieterse M, Bartkova J, Van Der Gulden H, Hiddingh S, Thanasoula M, Kulkarni A, Yang Q, et al. 53BP1 loss rescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers. Nat Struct Mol Biol 2010, 17:688-695. 10.1038/nsmb.1831, 2912507, 20453858.
67. Bunting SF, Callen E, Wong N, Chen HT, Polato F, Gunn A, Bothmer A, Feldhahn N, Fernandez-Capetillo O, Cao L, et al. 53BP1 inhibits homologous recombination in Brca1-deficient cells by blocking resection of DNA breaks. Cell 2010, 141:243-254. 10.1016/j.cell.2010.03.012, 2857570, 20362325.
68. Nik-Zainal S, Alexandrov LB, Wedge DC, Van Loo P, Greenman CD, Raine K, Jones D, Hinton J, Marshall J, Stebbings LA, et al. Mutational processes molding the genomes of 21 breast cancers. Cell 2012, 149:979-993. 10.1016/j.cell.2012.04.024, 3414841, 22608084.
69. Morales JC, Franco S, Murphy MM, Bassing CH, Mills KD, Adams MM, Walsh NC, Manis JP, Rassidakis GZ, Alt FW, Carpenter PB. 53BP1 and p53 synergize to suppress genomic instability and lymphomagenesis. Proc Natl Acad Sci USA 2006, 103:3310-3315. 10.1073/pnas.0511259103, 1413919, 16492765.
70. Chapman JR, Sossick AJ, Boulton SJ, Jackson SP. BRCA1-associated exclusion of 53BP1 from DNA damage sites underlies temporal control of DNA repair. J Cell Sci 2012, 125:3529-3534. 10.1242/jcs.105353, 22553214.
71. Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D, Lopez E, Kyle S, Meuth M, Curtin NJ, Helleday T. Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature 2005, 434:913-917. 10.1038/nature03443, 15829966.
72. Farmer H, McCabe N, Lord CJ, Tutt AN, Johnson DA, Richardson TB, Santarosa M, Dillon KJ, Hickson I, Knights C, et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 2005, 434:917-921. 10.1038/nature03445, 15829967.
73. Bunting SF, Callen E, Kozak ML, Kim JM, Wong N, Lopez-Contreras AJ, Ludwig T, Baer R, Faryabi RB, Malhowski A, et al. BRCA1 functions independently of homologous recombination in DNA interstrand crosslink repair. Mol Cell 2012, 46:125-135. 10.1016/j.molcel.2012.02.015, 22445484.
74. Dou H, Huang C, Van Nguyen T, Lu LS, Yeh ET. SUMOylation and de-SUMOylation in response to DNA damage. FEBS Lett 2011, 585:2891-2896. 10.1016/j.febslet.2011.04.002, 21486569.
75. Selvarajah J, Moumen A. Role of ubiquitination in the DNA damage response: proteomic analysis to identify new DNA-damage-induced ubiquitinated proteins. Biochem Soc Trans 2010, 38:87-91. 10.1042/BST0380087, 20074041.
76. Al-Hakim A, Escribano-Diaz C, Landry MC, O'Donnell L, Panier S, Szilard RK, Durocher D. The ubiquitous role of ubiquitin in the DNA damage response. DNA Repair (Amst) 2010, 9:1229-1240.
77. Stewart GS, Panier S, Townsend K, Al-Hakim AK, Kolas NK, Miller ES, Nakada S, Ylanko J, Olivarius S, Mendez M, et al. The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling cascade at sites of DNA damage. Cell 2009, 136:420-434. 10.1016/j.cell.2008.12.042, 19203578.
78. Shao Z, Davis AJ, Fattah KR, So S, Sun J, Lee KJ, Harrison L, Yang J, Chen DJ. Persistently bound Ku at DNA ends attenuates DNA end resection and homologous recombination. DNA Repair (Amst) 2012, 11:310-316.
79. Postow L, Ghenoiu C, Woo EM, Krutchinsky AN, Chait BT, Funabiki H. Ku80 removal from DNA through double strand break-induced ubiquitylation. J Cell Biol 2008, 182:467-479. 10.1083/jcb.200802146, 2500133, 18678709.
80. Feng L, Chen J. The E3 ligase RNF8 regulates KU80 removal and NHEJ repair. Nat Struct Mol Biol 2012, 19:201-206. 10.1038/nsmb.2211, 22266820.
81. Patel AG, Sarkaria JN, Kaufmann SH. Nonhomologous end joining drives poly(ADP-ribose) polymerase (PARP) inhibitor lethality in homologous recombination-deficient cells. Proc Natl Acad Sci USA 2011, 108:3406-3411. 10.1073/pnas.1013715108, 3044391, 21300883.