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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۴۲-۴۲
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عنوان انگلیسی Ps-52: A Unique Oct4 Interface and Its Role in Reprogramming to Induced Pluripotency
چکیده انگلیسی مقاله Objective: In 2006, the field of stem cell research was captivated by a possibility of epigenetic reprogramming of terminally differentiated cells to a pluripotent state by four factors - Oct4, Sox2, Klf4 and c-Myc. Hovewer, six years after this breakthrough, we still lack proper insight into this fascinating process on a molecular level. Among all four factors in the original cocktail, Oct4 evinces a special feature as it is impossible to replace this transcription factor by any other member of its protein family. But what makes the protein so unique? Materials and Methods: Purification and assembly of the Oct4POU:PORE complex; Crystallization and structure determination; Electrophoretic mobility shift assay; Reprogramming and rescue assay; Strep-tagged purification of proteins; Mass spectrometry; Yeast two hybrid screen; Genetic code expansion technology. Results: We solved the crystal structure of Oct4. This revealed a unique structured linker sequence of Oct4, which is not found in other POU class members. Moreover, this alpha-helical stretch is exposed to surface, creating an attractive interface regarding direct interactions with other components of the reprogramming machinery. A mutation screen along the linker proved several amino acids to play a crucial role in reprogramming. Based on current interactome studies, we conclude that the region is involved in recruiting key epigenetic players. Two different approaches have been chosen to determine the direct interaction partners of aforesaid Oct4 alpha helix. The results of a yeast two hybrid screen and in vivo crosslinking via the genetic code expansion technology will be presented. Conclusion: The linker region of Oct4 is involved in recruiting key epigenetic players during the reprogramming process.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/856
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