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Updated: May 9, 2026

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription
Published on: July 26, 2019
Structure and function of CarD, an essential mycobacterial transcription factor
Devendra B Srivastava1, Katherine Leon, Joseph Osmundson
1Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065, USA.
None:
CarD, an essential transcription regulator in Mycobacterium tuberculosis, directly interacts with the RNA polymerase (RNAP). We used a combination of in vivo and in vitro approaches to establish that CarD is a global regulator that stimulates the formation of RNAP-holoenzyme open promoter (RPo) complexes. We determined the X-ray crystal structure of Thermus thermophilus CarD, allowing us to generate a structural model of the CarD/RPo complex. On the basis of our structural and functional analyses, we propose that CarD functions by forming protein/protein and protein/DNA interactions that bridge the RNAP to the promoter DNA. CarD appears poised to interact with a DNA structure uniquely presented by the RPo: the splayed minor groove at the double-stranded/single-stranded DNA junction at the upstream edge of the transcription bubble. Thus, CarD uses an unusual mechanism for regulating transcription, sensing the DNA conformation where transcription bubble formation initiates.
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