Transcription Fidelity In Vitro

Range Table - link
Organism Budding yeast Saccharomyces cerevisiae
Reference Kireeva ML. et al., Transient reversal of RNA polymerase II active site closing controls fidelity of transcription elongation. Mol Cell. 2008 Jun 6 30(5):557-66. doi: 10.1016/j.molcel.2008.04.017. p.561 table 1PubMed ID18538654
Method P.561 caption beneath table 1: "TEC9 [elongation complex containing a 9 nt transcript] was obtained on the template 45C and chased with 5–500 mM CTP or 0.1–5 mM UTP. The error values for kpol and KD estimates were obtained from the hyperbolic fit as described in the Supplemental Experimental Procedures."
Comments P.560 left column bottom paragraph: "Comparisons of incorporation and misincorporation rates at physiological concentrations of NTPs by the WT and mutant Pol II variants provide only an estimation of the impact of the mutation on transcription fidelity. Because the mutation may affect the polymerization rates and the affinity to the nucleotide substrate, fidelity is calculated as the ratio of kpol/KD for the correct NTP to kpol/KD for the incorrect substrate (Wong et al., 1991). The apparent maximal polymerization rate, kpol, and the apparent dissociation constant, KD, were obtained for the WT and E1103G Pol II from the hyperbolic dependence of the reaction rates on the NTP concentration as described in the Supplemental Experimental Procedures. The resulting parameters are shown in Table 1."
Entered by Uri M
ID 110019