Studies on class III gene transcription.

In an attempt to understand the functional role of individual transcription factors, we have carried out S1 nuclease analyses of in vitro synthesized RNA using partially purified HeLa cell factors and truncated 5S and tDNA. The results indicate that faithful initiation of 5S gene transcription depends on the presence of IIIA, IIIB and IIIC in addition to RNA polymerase III. On the other hand, tDNA transcription initiation requires IIIB and IIIC and RNA polymerase III. In addition, we found that either 'A' block promoter or 'B' block alone can not serve as a template for the initiation of tDNA transcription. When an ultraviolet irradiated 5S DNA was used as a template, transcription is minimally affected at a lower dose suggesting that small differences in the helical structure may not have much effect on the transcription machinery. These results along with an in vitro attempt to reproduce Xenopus 5S RNA gene regulation are presented.