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The activity of transcription factor IIIC1 is impaired during differentiation of F9 cells

Wolfgang Meissner1, Renate Thomae, Klaus H Seifart

  • 1Institut für Molekularbiologie und Tumorforschung, Philipps Universität Marburg, Lahnstrasse 3, D-35037 Marburg, Germany. meissner@imt.uni-marburg.de

The Journal of Biological Chemistry
|December 14, 2001
PubMed

Insights

Mouse embryonal carcinoma cell differentiation involves reduced RNA polymerase III transcription. TFIIIC1 factor can restore this impaired transcription, especially early on, while TBP-TAF complex may also be limiting later.

Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Gene Regulation

Background:

  • Differentiation of mouse F9 embryonal carcinoma (EC) cells to parietal endoderm (PE) models early mouse embryo development.
  • This differentiation process is marked by a significant decrease in RNA polymerase III (pol III) transcription.

Purpose of the Study:

  • To investigate the specific pol III transcription factors that become limiting during EC cell differentiation.
  • To elucidate the role of transcription factor IIIC subunit 1 (TFIIIC1) and TATA-binding protein (TBP)-TAF complex in the down-regulation of pol III transcription.

Main Methods:

  • In vitro differentiation of mouse F9 EC cells.
  • Complementation assays using cell extracts and purified pol III transcription factors.
  • Partial purification of pol III transcription factors from differentiated and undifferentiated cells.
  • Analysis of type 2 and type 3 promoter transcription.

Main Results:

  • TFIIIC1 significantly restores impaired pol III transcription in early differentiation stages.
  • TBP-TAF complex (TFIIIBbeta) may become limiting at later stages but does not fully explain reduced transcription of type 2 promoters.
  • TFIIIC1 can reactivate U6 transcription (type 3 promoter), indicating TBP is not limiting.
  • TFIIIC1 activity is dramatically reduced in PE cell extracts, while TFIIIC2 activity is present in both EC and PE cells.

Conclusions:

  • TFIIIC1 is a key limiting factor for pol III transcription during early mouse EC cell differentiation.
  • Reduced TFIIIC1 activity contributes significantly to the overall down-regulation of pol III-dependent genes in parietal endoderm cells.
  • Other factors, in addition to TFIIIBbeta, are involved in the regulation of type 3 promoter transcription during differentiation.

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