A mutation in T7 RNA polymerase that facilitates promoter clearance Export

@article{citeulike:3380389, abstract = {10.1073/pnas.0407141102 Like multisubunit RNA polymerases (RNAPs), T7 RNAP frequently releases its transcript over the initial 8-12 transcribed nucleotides, when it still contacts the promoter. This abortive cycling, which is most prominent with initial sequences that deviate from those of T7 late genes, eventually compromises productive transcription. Starting from an situation where transcription of a target gene by T7 RNAP is virtually abolished because of extensive abortive cycling, we have selected a mutation in RNAP that restores target gene expression. , this mutation (P266L) weakens promoter binding but markedly reduces abortive cycling over a variety of initial sequences by stabilizing the transcription complex at nucleotides 5-8. Other substitutions of P266 have similar effects. X-ray data show that during the transition from initial to elongation complex, the N-terminal region undergoes a major structural switch of which P266 constitutes one of the hinges. How the mutation might facilitate this switch is tentatively discussed. On the practical side, the mutation can significantly improve transcription, particularly from templates carrying unfavorable initial sequences.}, author = {Guillerez, Jean and Lopez, Pascal J. and Proux, Florence and Launay, H\'{e}l\`{e}ne and Dreyfus, Marc}, citeulike-article-id = {3380389}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0407141102}, citeulike-linkout-1 = {http://www.pnas.org/content/102/17/5958.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/102/17/5958.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/15831591}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=15831591}, day = {26}, doi = {10.1073/pnas.0407141102}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {rna-production}, month = {April}, number = {17}, pages = {5958--5963}, posted-at = {2008-10-06 17:36:00}, priority = {3}, title = {A mutation in T7 RNA polymerase that facilitates promoter clearance}, url = {http://dx.doi.org/10.1073/pnas.0407141102}, volume = {102}, year = {2005} }

TY - JOUR ID - citeulike:3380389 L3 - citeulike-article-id:3380389 N2 - 10.1073/pnas.0407141102 Like multisubunit RNA polymerases (RNAPs), T7 RNAP frequently releases its transcript over the initial 8-12 transcribed nucleotides, when it still contacts the promoter. This abortive cycling, which is most prominent with initial sequences that deviate from those of T7 late genes, eventually compromises productive transcription. Starting from an situation where transcription of a target gene by T7 RNAP is virtually abolished because of extensive abortive cycling, we have selected a mutation in RNAP that restores target gene expression. , this mutation (P266L) weakens promoter binding but markedly reduces abortive cycling over a variety of initial sequences by stabilizing the transcription complex at nucleotides 5-8. Other substitutions of P266 have similar effects. X-ray data show that during the transition from initial to elongation complex, the N-terminal region undergoes a major structural switch of which P266 constitutes one of the hinges. How the mutation might facilitate this switch is tentatively discussed. On the practical side, the mutation can significantly improve transcription, particularly from templates carrying unfavorable initial sequences. IS - 17 JF - Proceedings of the National Academy of Sciences of the United States of America EP - 5963 TI - A mutation in T7 RNA polymerase that facilitates promoter clearance VL - 102 SP - 5958 KW - rna-production AU - Guillerez, Jean AU - Lopez, Pascal AU - Proux, Florence AU - Launay, Hélène AU - Dreyfus, Marc PY - 2005/04/26/ UR - http://dx.doi.org/10.1073/pnas.0407141102 ER -