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Consistently, the co-transfection assay of COP1-binding defective DET1 mutants including R488A/H485G, R488A/H485G/R306G/S312G and R488A/H485G/E380A/T382A with COP1 and Ube2e2 demonstrated that the reduction of COP1-DET1 interaction resulted in the decreased amount of Ube2e2 co-purification, suggesting that DET1-mediated proximity is not sufficient for COP1 and Ube2e2 to directly interact (Fig