Related Experiment Video
Updated: Jun 10, 2026

Heterokaryon Technique for Analysis of Cell Type-specific Localization
Published on: March 11, 2011
Interaction between nuclear localization signal-retinoic acid receptor alpha and Ubiquilin 1
Dan Zhu1, Chong Wang, Beizhong Liu
1Key Laboratory of Laboratory Medicine Diagnostics, Ministry of Education of People's Repulic of China, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China.
Objective:
To identify the interaction between nuclear localization signal-retinoic acid receptor alpha (NLS-RARalpha) and Ubiquilin 1(UBQLN1).
Methods:
The recombination expression plasmids pGBKT7-NLS-RAR and pACT2-UBQLN1, which expressed bait-protein NLS-RARalpha and target protein UBQLN1 respectively, were cotransformated into yeast AH109. The interaction of the expression plasmids in the living cells was investigated by yeast two-hybrid assay. HA-tagged fusion protein (pCMV-HA-NLS-RARalpha) expression vector and Myc-tagged fusion protein (pCMV-Myc-JTV1) expression vector were constructed, identified, and cotransfected respectively into human embryo kidney 293 cells(HEK293). The interaction was detected by co-immunoprecipitation.
Results:
Blue clones were found on yeast AH109 plate cotransformated with pGBKT7-NLS-RARalpha and pACT2-UBQLN1. Double restriction enzyme digestion showed that pCMV-HA-NLS-RARalpha and pCMV-Myc-JTV1 were successfully constructed. Then HA-NLS-RAR protein was immunoprecipitated by anti-HA polyclonal antibody and the expression of Myc-UBQLN1 was tested by Western blot with anti-Myc monoclonal antibody from immunoprecipitated complex.
Conclusion:
The interaction between NLS-RARalpha and UBQLN1 can be verified by yeast two-hybrid assay and co-immunoprecipitation.
Related Concept Videos
Nuclear Localization Signals and Import
Regulation of Nuclear Protein Sorting
Nuclear Protein Sorting
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
Receptor Downregulation in MVBs
The EGFR can initiate signaling pathways that lead to cell proliferation, migration, and differentiation. Overexpression of EGFR stimulates cells to proliferate. Excessive EGFR activation may...
Directing Proteins to the Rough Endoplasmic Reticulum
Nuclear Export
NES are of three types- the canonical 10-residue long leucine-rich signal and other...

