Structural basis for the bacterial transcription-repair coupling factor/RNA polymerase interaction.
The transcription-repair coupling factor (TRCF, the product of the mfd gene) is a widely conserved bacterial protein that mediates transcription-coupled DNA repair. TRCF uses its ATP-dependent DNA translocase activity to remove transcription complexes stalled at sites of DNA damage, and stimulates repair by recruiting components of the nucleotide excision repair pathway to the site. A protein/protein interaction between TRCF and the β-subunit of RNA polymerase (RNAP) is essential for TRCF function. CarD (also called CdnL), an essential regulator of rRNA transcription in Mycobacterium tuberculosis, shares a homologous RNAP interacting domain with TRCF and also interacts with the RNAP β-subunit. We determined the 2.9-Å resolution X-ray crystal structure of the RNAP interacting domain of TRCF complexed with the RNAP-β1 domain, which harbors the TRCF interaction determinants. The structure reveals details of the TRCF/RNAP protein/protein interface, providing a basis for the design and interpretation of experiments probing TRCF, and by homology CarD, function and interactions with the RNAP.
Westblade
Lars F
LF
Laboratory of Molecular Biophysics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
Campbell
Elizabeth A
EA
Pukhrambam
Chirangini
C
Padovan
Julio C
JC
Nickels
Bryce E
BE
Lamour
Valerie
V
Darst
Seth A
SA
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018360:Crystallography, X-Ray; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D054730:Protein Interaction Domains and Motifs; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D014157:Transcription Factors; C072103:transcription repair coupling factor protein, Bacteria; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit