Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Effects of a Phosphomimetic Mutant of RAP80 on Linear Polyubiquitin Binding Probed by Calorimetric Analysis

  • Thach, Thanh Trung (Department of Biological Sciences, Sungkyunkwan University) ;
  • Jee, Jun-Goo (Research Institute of Pharmaceutical Sciences, College of Pharmacy, Kyungpook National University) ;
  • Lee, Sang-Ho (Department of Biological Sciences, Sungkyunkwan University)
  • Received : 2012.01.12
  • Accepted : 2012.01.20
  • Published : 2012.04.20
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Abstract

RAP80 plays a key role in DNA damage responses by recognizing K63-linked polyubiquitin moieties through its two ubiquitin-interacting motif (UIM) domains. The linker between the two UIMs possesses a phosphorylation site, but the relationship between phosphorylation and polyubiquitin recognition remains elusive. We investigated the interaction between a phosphorylation-mimic RAP80 mutant S101E and linear polyubiquitins, structurally equivalent to the K63-linked ones, using isothermal titration calorimetry (ITC). ITC analysis revealed differential binding affinities for linear tetraubiquitin by otherwise equivalent UIMs in S101E. Mutational analysis supported such differential polyubiquitin recognition by S101E. Our results suggest a potential crosstalk between polyubiquitin recognition and phosphorylation in RAP80.

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