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Mahdi et al. (2003) EMBO J. 22, 724.Chambers et al. (2003) Nucleic Acids Res. 31, 6409.
Rescue• Transcript cleavage (stimulated by GreA/GreB)• Slide forward
Park et al. (2002) Cell 109, 757
Requirements for TRCF activity on the elongation complex
RNAPRNAPRNAPRNAPRNAPRNAPRNAPRNAPRNAPRNAPRNAP
RNAP
translocation domains
wedge domain
RNAP binding domain
RecG
Singleton et al. (2001) Cell 107, 79
TRCF
Park et al. (2002) Cell 109, 757
Alexandra Deaconescu
Collaborators
Anna L. Chambers, Abigail J. Smith, Nigel J. Savery (University of Bristol, Bristol, UK)
Bryce E. Nickels, Ann Hochschild (Harvard Medical School)
Data collection and model refinement statistics for apo Mfd
Space group: C2; 2 molecules per asymmetric unit SeMet1 SeMet2Cell dimensions (Å): a=151.7 b=162.2 c=162.1 a=151.9 b=162.0 c= 161.7 β= 104.9° β= 105.1°Wavelengt (h Å) 0.98166 0.97899Resolut ion( ) 40-4.0 40Å -3.2Reflecti onsmeasure 228,334 300,181d(unique) 62,430 122,388Completene ss(%) 99.6 99.9(i n oute r shell) 100.0 99.9Mea n I/σ(I) 10.9 12.4(i n oute r shell) 2.9 2.5Rs ym
a(%) 12.9 0.08(i n oute r shell) 50.2 37.7
S AD Figu re o f Meri 0.17tRefinement dat ase : t SeM 2 (|ET F| >0)R-facto r 23.4% R-fre efact 29.5%orR. .ms. d bondle ngth( )Å : 0.011R. .ms. d bondangle s (°): 1.49
aRsym=Σ|I-<I>|/Σ , I wher e =I obse rvedintensit , <y I>=avera geintensi ty o f multipleobservati ons o f symmetr -y relat ed reflectionsbRcryst=Σ|Fo - FcI/Σ|Fo|, whe reFo andFc ar e obser ved andcalculate dstructur efactors,respectively
R/Rfree = 0.25/0.29 (3.2 Å resolution)APS NE-CAT-8BM
APS SBC-19IDDeaconescu et al. (2006)
Cell 124, 507.
QuickTime™ and aCompact Video decompressorare needed to see this picture.
Singleton et al. (2001) Cell 107, 79Deaconescu et al. (2006) Cell 124, 507.
UvrB homology module, UvrA recruitment, and the role of D7
Truglio et al. (2002) EMBO
J. 23, 2498
• necessary for RNAP binding (Selby & Sancar, 1995)• binds RNAP β subunit 19-142 (Park et al., 2002)
DNA-binding and ATPase activity of TRCF-L499R
mutant intact
Deaconescu et al. (2006) Cell 124, 507.
Deaconescu et al. (2006) Cell 124, 507.
Park et al. (2002) Cell 109, 757
A. L. Chambers, A.J. Smith, N. J. Savery
B. Nickels, A. HochschildLeu499
Park et al. (2002) Cell 109, 757Smith & Savery (2005) Nucleic Acids Res. 33, 755
Dürr et al. (2005) Cell 121, 363
• TRCF - compact arrangement of structural domains linked by long, flexible linkers - appears ‘primed’ for large scale conformational changes (translocation module, UvrA binding surface).
? Nature and timing of TRCF conformational changes during the steps of TRCF-mediated transcription-coupled repair reaction? (recognition of a stalled RNAP; forward translocation of the RNAP to the transcription block; RNAP release and transcript termination; recruitment of the NER machinery to the site)
? Details of protein/protein interactions involving TRCF? (TRCF/RNAP and TRCF/NER machinery)