This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Culotta, V C Articles by Sollner-Webb, B Search for Related Content PubMed PubMed Citation Articles by Culotta, V C Articles by Sollner-Webb, B

 Previous Article  |  Next Article 

Mol Cell Biol. 1985 July; 5(7): 1582-1590

Eucaryotic transcription complexes are specifically associated in large sedimentable structures: rapid isolation of polymerase I, II, and III transcription factors.

ABSTRACT

RNA synthesis in eucaryotes takes place on template molecules that are activated by stably associating with limiting transcription factors. In this paper we demonstrate that such stable transcription complexes can be specifically sedimented from in vitro transcription reaction mixtures by mild centrifugation. This occurs with stable complexes of genes transcribed by all three classes of eucaryotic RNA polymerase and with S-100 as well as whole-cell extracts. However, the transcriptional capacity of the isolated complex differs for the three polymerase classes. The pelleted ribosomal DNA (polymerase I) complex contains all the factors necessary for transcription, each purified 25- to 50-fold, whereas the pelleted adenovirus major late promoter (polymerase II) complex lacks a factor that remains in the supernatant. In the case of 5S DNA (polymerase III), a necessary factor associates slowly with the sedimentable complex. Notably, the interactions responsible for this rapid sedimentation are specific for DNA molecules in stable complexes, suggesting that the in vitro sedimentable complex mirrors the in vivo structural organization of active genes.

This article has been cited by other articles:

• Cook, P. R. (1999). The Organization of Replication and Transcription. Science 284: 1790-1795 [Abstract] [Full Text]  
• Riggs, D. L., Peterson, C. L., Wickham, J. Q., Miller, L. M., Clarke, E. M., Crowell, J. A., Sergere, J.-C. (1995). Characterization of the Components of Reconstituted Saccharomyces cerevisiae RNA Polymerase I Transcription Complexes. J. Biol. Chem. 270: 6205-6210 [Abstract] [Full Text]