Figure 3
From: The E3 ubiquitin ligase TRIP12 participates in cell cycle progression and chromosome stability
TRIP12 is associated with chromatin through a N-terminal intrinsically disordered region. (A) Prediction of intrinsically disordered and ordered regions in TRIP12 protein sequence (Protein accession number NP_004229.1) was determined using IUPred, GlobPlot and DisEMBL software. aa stands for amino acids. (B) GFP-fusion protein localization in metaphasic HelaS3 cells transfected with TRIP12 (1-1992), (1-1552), (1-445), (446-1992) and (1553-1992)-GFP constructs was visualized by immunofluorescence using an anti-GFP antibody. H2B-GFP and CAT-GFP constructs were used as controls. Nuclei were counterstained with DAPI. IDR stands for Intrinsically Disordered Region, NLS for Nuclear Localization Signal, ARM for β-Armadillo domain, WWE for tryptophan and glutamate enriched domain and HECT for Homologous to the E6-AP Carboxyl Terminus. Numbers indicate amino acids (aa) position. (C) GFP-fusion protein localization in interphasic HelaS3 cells transfected with TRIP12 (1-1992), (1-445), (446-1000), (1001-1552) and (1553-1992)-GFP constructs was analyzed by immunofluorescence using an anti-GFP antibody. H2B-GFP and CAT-GFP constructs were used as controls. Nuclei were counterstained with DAPI. IDR stands for Intrinsically Disordered Region, ARM for β-Armadillo domain, WWE for tryptophan and glutamate enriched domain and HECT for Homologous to the E6-AP Carboxyl Terminus. Numbers indicate amino acids (aa) position. (D) Interaction of increasing amounts of GST-TRIP12-ARM-WWE(446-1000) and GST-IDR(1-325) with naked plasmid DNA visualized after migration on agarose gel.
