CarD uses a minor groove wedge mechanism to stabilize the RNA polymerase open promoter complex

Bae, Brian; Chen, James; Davis, Elizabeth; Leon, Katherine; Darst, Seth A.; Campbell, Elizabeth A.

doi:10.7554/eLife.08505

CarD uses a minor groove wedge mechanism to stabilize the RNA polymerase open promoter complex

Journal Article · Tue Sep 08 00:00:00 EDT 2015 · eLife

DOI:https://doi.org/10.7554/eLife.08505· OSTI ID:1233324

Bae, Brian ^[1]; Chen, James ^[1]; Davis, Elizabeth ^[1]; Leon, Katherine ^[1]; Darst, Seth A. ^[1]; Campbell, Elizabeth A. ^[1]

Rockefeller Univ., New York, NY (United States)

A key point to regulate gene expression is at transcription initiation, and activators play a major role. CarD, an essential activator in Mycobacterium tuberculosis, is found in many bacteria, including Thermus species, but absent in Escherichia coli. To delineate the molecular mechanism of CarD, we determined crystal structures of Thermus transcription initiation complexes containing CarD. The structures show CarD interacts with the unique DNA topology presented by the upstream double-stranded/single-stranded DNA junction of the transcription bubble. We confirm that our structures correspond to functional activation complexes, and extend our understanding of the role of a conserved CarD Trp residue that serves as a minor groove wedge, preventing collapse of the transcription bubble to stabilize the transcription initiation complex. Unlike E. coli RNAP, many bacterial RNAPs form unstable promoter complexes, explaining the need for CarD.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: National Center for Research Resources (NCRR); National Institute of Biomedical Imaging and Bioengineering (NIBIB); National Institute of General Medical Sciences (NIGMS)

OSTI ID:: 1233324

Alternate ID(s):: OSTI ID: 1354289

Journal Information:: eLife, Journal Name: eLife Journal Issue: 2015 Vol. 4; ISSN 2050-084X

Publisher:: eLife Sciences Publications, Ltd.Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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