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[BK21 Plus Seminar]
▶Subject: Direct analysis of phosphorylation sites on the Rpb1 C-terminal Domain of RNA polymerase II utilizing mass spectrometry
▶Speaker: HYUNSUK SUH, PH.D.(Harvard Medical School)
▶Date: 4:30 PM/August. 23(Tue.)/2016
▶Place: Auditorium(1F), Postech Biotech Center
Dynamic interactions between RNA polymerase II (RNApII) and various mRNA processing and chromatin modifying enzymes are mediated by the changing phosphorylation pattern on the C-terminal domain (CTD) of polymerase subunit Rpb1 during different stages of transcription. Since the first report of CTD three decades ago, phosphorylations within the repetitive heptamer sequence (Y1-S2-P3-T4-S5-P6-S7) of CTD have primarily been defined using antibodies, but these do not distinguish different repeats or allow comparative quantitation. To address these questions, a CTD modified for mass spectrometry (msCTD) was utilized and revealed the spatial pattern and relative abundance of phosphorylations along the domain. MS analyses show that Ser5-P and Ser2-P occur throughout the length of CTD and are far more abundant than other phosphorylation sites. msCTD extracted from cells mutated in several CTD kinases or phosphatases showed the expected changes in phosphorylation. Furthermore, msCTD associated with capping enzyme was enriched for Ser5-P while that bound to the transcription termination factor Rtt103 had higher levels of Ser2-P. These results suggest a relatively sparse and simple "CTD code". Further studies based on this approach are anticipated to contribute to elucidating how RNApII and regulatory factors communicate during transcription.
▶Inquiry: Prof. Lee, Yoontae (279-2354)
* This seminar will be given in English.
please refrain from taking photos during seminars. *