Journal
JOURNAL OF GENETICS AND GENOMICS
Volume 35, Issue 12, Pages 723-728Publisher
SCIENCE PRESS
DOI: 10.1016/S1673-8527(08)60227-1
Keywords
Oct4; karyopherin-alpha 2; yeast two-hybrid; nuclear localization signal
Funding
- Shanghai Science & Technology Developmental Foundations [06dj14001]
- National High Technology Research and Development Program of China [2006AA02Z197, 2006CB943901, 2007CB947904]
- Shanghai Jiao Tong University School of Medicine and Shanghai Institutes for Biological Sciences, CAS
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The POU domain transcription factor Oct4 is a master regulator in maintaining self-renewal and pluripotency of embryonic stem (ES) cells. To further explore the functional network of Oct4, the yeast two-hybrid system was used to search for Oct4 interacting proteins. PH domain (containing POU domain and homeodomain) of human OCT4 was used as a bait. From the human testis cDNA library, we identified a strong interaction between OCT4 and karyopherin-alpha 2 (KPNA-2). KPNA2 is involved in active nuclear import of proteins. This finding was confirmed by glutathione S-transferase pull-down and co-immunoprecipitation assays. The interaction between OCT4 and KPNA-2 was further mapped to multiple regions of the two proteins. In addition, we studied nuclear localization signal (NLS) of mouse Oct4 and demonstrated that it is essential for Oct4 nuclear localization. Thus, our data suggest that Oct4 nuclear localization may be mediated by its interaction with KPNA-2.
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