Nucleic Acids Research, 1993, Vol. 21, No. 18 4180-4186
© 1993
MOLECULAR BIOLOGY |
A TBP-containing multiprotein complex (TIF-IB) mediates transcription specificity of murine RNA polymerase I
Institute of cell and Turmor Biology, German Cancer Research Center D-69120 Heidelberg, Germay 1Institut de Chimie Biologique, Faculteè de Meèdecine 11 rue Humman, 67085 Strasbourg Ceèdex, France
*To whom correspondence should be addressed
Received July 6, 1993. Revised August 2, 1993. Accepted August 2, 1993.
TIF-IB is a transcription factor which interacts with the mouse ribosomal gene promoter and nucleates the formation of an initiation complex containing RNA polymerase I (Pol I). We have purified this factor to near homogeneity and demonstrate that TIF-IB is a large complex (>200 kDa) which contains several polypeptides. One of the subunits present In this protein complex Is the TATA–blnding protein (TBP) as revealed by copurlflcation of TIF–IB activity and TBP over different chromatographlc steps including Immunoafflnity purification. In addition to TBP, three tightly associated proteins (TAFs-l) with apparent molecular weights of 95, 68, and 48 kDa are contained In this multimeric complex. This subunit composition Is similar but not Identical to the analogous human factor SL1. Depletion of TBP from TIF-IB-containlng fractions by Immunoprecipltation eliminates TIF-IB activity. Neither TBP alone nor fractions containing other TBP complexes are capable of substituting for TIF-IB activity. Therefore, TIF-IB is a unique complex with Pol l-speciflc TAFs distinct from other TBPcontaining complexes. The Identification of TBP as an integral part of the murine rDNA promoter-specific transcription Initiation factor extends the previously noted similarity of transcriptional initiation by the three nuclear RNA polymerases and underscores the Importance of TAFs in determining promoter specificity.
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