Open AccessReconstitution of Pol II (G) responsive form of the human Mediator complex
Author(s) -
Murat Cevher
Publication year - 2021
Publication title -
turkish journal of biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.323
H-Index - 38
eISSN - 1303-6092
pISSN - 1300-0152
DOI - 10.3906/biy-2009-12
Subject(s) - mediator , rna polymerase ii , biology , protein subunit , transcription factor ii d , rna polymerase iii , microbiology and biotechnology , transcription (linguistics) , activator (genetics) , gene , rna polymerase , gene expression , genetics , promoter , rna , linguistics , philosophy
RNA polymerase II (Pol II) is a 12 subunit protein complex from yeast to human that is required for gene expression. Gdown1 containing Pol II [Pol II (G)] is a special form of Pol II that is catalytically inactive and heavily depends on the 30-subunit Mediator complex for its activator and basal dependent function in vitro. Here we report for the first time, the identification and the generation of a 15-subunit human Mediator complex via the novel multibac baculovirus expression system that is fully responsive to Pol II (G). Our results show complete recovery of Pol II (G) dependent transcription both with full 30-subunit Mediator and also with 15-subunit recombinant Mediator that we synthesized. Moreover, we also show that the recombinant Mediator interacts with Pol II (G) as well. These results enlighten us towards understanding how a certain population of Pol II that is involved in selected gene regulation is activated by Mediator complex.