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J. Biol. Chem., Vol. 275, Issue 29, 22104-22113, July 21, 2000

This Article Full Text Full Text (PDF) All Versions of this Article: 275/29/22104    most recent M002253200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chaney, M. Articles by Buck, M. Search for Related Content PubMed PubMed Citation Articles by Chaney, M. Articles by Buck, M. Social Bookmarking                      What's this?

Sequences within the DNA Cross-linking Patch of ⁵⁴ Involved in Promoter Recognition,  Isomerization, and Open Complex Formation^*

Matthew Chaney, Melinda Pitt, and Martin Buck^§

From the Department of Biology, Imperial College of Science, Technology, and Medicine, London SW7 2AZ, United Kingdom

The bacterial RNA polymerase holoenzyme containing the ⁵⁴ subunit functions in enhancer-dependent transcription. Mutagenesis has been used to probe the function of a sequence in the ⁵⁴ DNA binding domain that includes residues that cross-link to promoter DNA. Several activities of the  and holoenzyme are shown to depend on the cross-linking patch. The patch contributes to promoter binding by ⁵⁴, and holoenzyme and is involved in activator-dependent  isomerization. As part of the ⁵⁴-holoenzyme, some residues in the patch limit basal transcription. Other cross-linking patch sequences appear to limit activator-dependent open complex formation. Deletion of 19 residues adjacent to the cross-linking patch resulted in a holoenzyme unable to respond to activator but capable of activator-independent (bypass) transcription in vitro. Overall results are consistent with the cross-linking patch directing interactions to the 12 promoter region to set basal and activated levels of transcription.

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