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1 Laboratory of Biochemistry and Molecular Biology, The
Rockefeller University, New York, New York 10021 USA;
2 Stuyvesant High School, New York, New York 10282 USA
Transcription by RNA polymerase III (Pol III) requires multiple
general initiation factors that, in isolated form, assemble onto the
promoter in an ordered fashion. Here, it is shown that all components
required for transcription of the VA1 and tRNA genes, including TFIIIB,
TFIIIC, and RNA Pol III, can be coimmunopurified from a HeLa cell line
that constantly expresses a FLAG epitope-tagged subunit of human RNA
Pol III. This finding of an RNA Pol III "holoenzyme" suggests
similarities between transcription initiation by RNA Pol II and RNA Pol
III and has led to the identification of a novel general initiation
factor (TDF, translation dependent
factor) that is present within the holoenzyme. TDF is
selectively inactivated during protein synthesis inhibition by
cycloheximide and at a late stage of adenovirus infection, thus
accounting for the loss of RNA Pol III-mediated transcription of the
tRNA and VA RNA genes under these conditions. On the basis of these
observations, possible mechanisms for the global regulation of
transcription by RNA Pol III and for disassembly of RNA Pol III
initiation complexes are proposed.
[Key Words: Transcription; holoenzyme; RNA polymerase III]
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