RNF8 E3 Ubiquitin Ligase Stimulates Ubc13 E2 Conjugating Activity That Is Essential for DNA Double Strand Break Signaling and BRCA1 Tumor Suppressor Recruitment
In: The Journal of biological chemistry, vol 291, iss 18 Hodge, CD; Ismail, IH; Edwards, RA; Hura, GL; Xiao, AT; Tainer, JA; et al.(2016). RNF8 E3 ubiquitin ligase stimulates Ubc13 E2 conjugating activity that is essential for DNA double strand break signaling and BRCA1 tumor suppressor recruitment. Journal of Biological Chemistry, 291(18), 9396-9410. doi: 10.1074/jbc.M116.715698. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/3sg7k2t8; (2016-04-01)
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Zugriff:
DNA double strand break (DSB) responses depend on the sequential actions of the E3 ubiquitin ligases RNF8 and RNF168 plus E2 ubiquitin-conjugating enzyme Ubc13 to specifically generate histone Lys-63-linked ubiquitin chains in DSB signaling. Here, we defined the activated RNF8-Ubc13∼ubiquitin complex by x-ray crystallography and its functional solution conformations by x-ray scattering, as tested by separation-of-function mutations imaged in cells by immunofluorescence. The collective results show that the RING E3 RNF8 targets E2 Ubc13 to DSB sites and plays a critical role in damage signaling by stimulating polyubiquitination through modulating conformations of ubiquitin covalently linked to the Ubc13 active site. Structure-guided separation-of-function mutations show that the RNF8 E2 stimulating activity is essential for DSB signaling in mammalian cells and is necessary for downstream recruitment of 53BP1 and BRCA1. Chromatin-targeted RNF168 rescues 53BP1 recruitment involved in non-homologous end joining but not BRCA1 recruitment for homologous recombination. These findings suggest an allosteric approach to targeting the ubiquitin-docking cleft at the E2-E3 interface for possible interventions in cancer and chronic inflammation, and moreover, they establish an independent RNF8 role in BRCA1 recruitment.
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RNF8 E3 Ubiquitin Ligase Stimulates Ubc13 E2 Conjugating Activity That Is Essential for DNA Double Strand Break Signaling and BRCA1 Tumor Suppressor Recruitment
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Autor/in / Beteiligte Person: | Hura, Greg L. ; Ismail Hassan Ismail ; Tainer, John A. ; Edwards, Ross A. ; Xiao, Andrew T. ; Hendzel, Michael J. ; Hodge, Curtis D. ; J. N. Mark Glover |
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Quelle: | The Journal of biological chemistry, vol 291, iss 18 Hodge, CD; Ismail, IH; Edwards, RA; Hura, GL; Xiao, AT; Tainer, JA; et al.(2016). RNF8 E3 ubiquitin ligase stimulates Ubc13 E2 conjugating activity that is essential for DNA double strand break signaling and BRCA1 tumor suppressor recruitment. Journal of Biological Chemistry, 291(18), 9396-9410. doi: 10.1074/jbc.M116.715698. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/3sg7k2t8; (2016-04-01) |
Veröffentlichung: | eScholarship, University of California, 2016 |
Medientyp: | unknown |
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