Fig. 1
UBN1 and UBN2 are responsible for the specific recognition and direct binding of H3.3. a UBN1 specifically recognizes H3.3. Interaction between HIRA, UBN1, Cabin1, Asf1a, and H3.1, H3.3 is analyzed by Western blot analysis of anti-Flag immunoprecipitates. b HIRA is dispensable for the interaction between UBN1 and H3.3. The interaction between UBN1 and H3.3 is analyzed by immunoprecipitation in wild type (WT) or HIRA knockout (HIRA KO) HEK293T cells as illustrated. c HIRA enhances the interaction between UBN1 and H3.3. The effect of HIRA and Cabin1 on the interaction between UBN1 and H3.3 is analyzed by Western blot analysis of anti-Flag immunoprecipitates. d Both UBN1 and UBN2 specifically interact with H3.3. Interaction between UBN1, UBN2, and H3.1, H3.3 in the presence of HIRA is analyzed by Western blot analysis of anti-Flag immunoprecipitates. e The direct interaction between UBN1 (aa1-213), UBN2 (aa109-291), and (H3.1-H4)2, (H3.3-H4)2 tetramers is analyzed by GST pull-down followed by Coomassie staining. Input corresponds to 10% proteins. * indicates nonspecific band in GST-UBN2-(aa109-291) sample