A test of the model that RNA polymerase III transcription is regulated by selective induction of the 110 kDa subunit of TFIIIC.
TFIIIC is a RNA polymerase (pol) III-specific DNA-binding factor that is required for transcription of tRNA and 5S rRNA genes. Active human TFIIIC consists of five subunits. However, an inactive form has also been isolated that lacks one of the five subunits, called TFIIIC110. A model was proposed in which pol III transcription might be regulated by the specific induction of TFIIIC110, allowing formation of active TFIIIC from the inactive form. We have tested this model by transient transfection of HeLa and HEK293 cells with a vector expressing TFIIIC110. We have also made stably transfected HeLa cell lines that carry a doxycycline-inducible version of the cDNA for TFIIIC110. We show that the induced TFIIIC110 enters the nucleus, binds other TFIIIC subunits and is recruited to tRNA and 5S rRNA genes in vivo. However, little or no effect is seen on the expression of pol III transcripts. The data argue against the model that pol III transcription can be effectively modulated through the specific induction of TFIIIC110.
Innes
Fiona
F
Institute of Biomedical and Life Sciences, Division of Biochemistry and Molecular Biology, University of Glasgow, Glasgow G12 8QQ, UK.
Ramsbottom
Ben
B
White
Robert J
RJ
D002460:Cell Line; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D008957:Models, Genetic; D021122:Protein Subunits / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012320:RNA Polymerase III / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000096:biosynthesis; D012343:RNA, Transfer / Q000096:biosynthesis; D035361:Transcription Factor TFIIIB / Q000378:metabolism; D019474:Transcription Factors, TFIII / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D000098832:Transcription Factor TFIIIC
D021122:Protein Subunits; D012341:RNA, Ribosomal, 5S; D035361:Transcription Factor TFIIIB; D019474:Transcription Factors, TFIII; D000098832:Transcription Factor TFIIIC; D012343:RNA, Transfer; D012320:RNA Polymerase III