TY - GEN
T1 - A strategy for dissecting the kinetics of transcription repression mechanisms
AU - Palma, Cristina S.D.
AU - Startceva, Sofia
AU - Neeli-Venkata, Ramakanth
AU - Zare, Marzieh
AU - Goncalves, Nadia S.M.
AU - Fonseca, Jose M.
AU - Oliveira, Samuel M.D.
AU - Ribeiro, Andre S.
N1 - jufoid=58152
INT=tut-bmt,"Zare, Marzieh"
PY - 2018
Y1 - 2018
N2 - Promoters in Escherichia coli include an ‘OFF’ state, during which transcription is halted. Here, we propose a novel empirical method for assessing the time-length spent by promoters in this state. It relies on direct measurements of RNA production kinetics at the single molecule level at different induction levels, followed by an estimation of the RNA production rate under infinite induction, which is then compared to this rate under real, maximum induction. We apply it to the LacO3O1 promoter and infer that, under full induction, on average, 15% of the time between successful transcription events is spent in the OFF state. We verify this result by comparing the kinetics of a mutant strain lacking repressor molecules with that of the inferred rate under infinite induction. We expect this strategy of dissecting the kinetics of transcription repression to be applicable to a wide number of promoters in E. coli.
AB - Promoters in Escherichia coli include an ‘OFF’ state, during which transcription is halted. Here, we propose a novel empirical method for assessing the time-length spent by promoters in this state. It relies on direct measurements of RNA production kinetics at the single molecule level at different induction levels, followed by an estimation of the RNA production rate under infinite induction, which is then compared to this rate under real, maximum induction. We apply it to the LacO3O1 promoter and infer that, under full induction, on average, 15% of the time between successful transcription events is spent in the OFF state. We verify this result by comparing the kinetics of a mutant strain lacking repressor molecules with that of the inferred rate under infinite induction. We expect this strategy of dissecting the kinetics of transcription repression to be applicable to a wide number of promoters in E. coli.
KW - Induction
KW - OFF state
KW - Transcription
KW - τ plot
U2 - 10.1007/978-981-10-5122-7_274
DO - 10.1007/978-981-10-5122-7_274
M3 - Conference contribution
AN - SCOPUS:85021725472
SN - 9789811051210
T3 - IFMBE Proceedings
SP - 1097
EP - 1100
BT - EMBEC and NBC 2017 - Joint Conference of the European Medical and Biological Engineering Conference EMBEC 2017 and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics, NBC 2017
PB - Springer Verlag
T2 - Joint Conference of the European Medical and Biological Engineering Conference (EMBEC) and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics (NBC)
Y2 - 1 January 1900
ER -