Live imaging of nascent RNA dynamics reveals distinct types of transcriptional pulse regulation. / Muramoto, Tetsuya; Cannon, Danielle; Gierlinski, Marek; Corrigan, Adam; Barton, Geoffrey J.; Chubb, Jonathan R.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, No. 19, 2012, p. 7350-7355.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Live imaging of nascent RNA dynamics reveals distinct types of transcriptional pulse regulation
A1 - Muramoto,Tetsuya
A1 - Cannon,Danielle
A1 - Gierlinski,Marek
A1 - Corrigan,Adam
A1 - Barton,Geoffrey J.
A1 - Chubb,Jonathan R.
AU - Muramoto,Tetsuya
AU - Cannon,Danielle
AU - Gierlinski,Marek
AU - Corrigan,Adam
AU - Barton,Geoffrey J.
AU - Chubb,Jonathan R.
PY - 2012
Y1 - 2012
N2 - <p>Transcription of genes can be discontinuous, occurring in pulses or bursts. It is not clear how properties of transcriptional pulses vary between different genes. We compared the pulsing of five housekeeping and five developmentally induced genes by direct imaging of single gene transcriptional events in individual living Dictyostelium cells. Each gene displayed its own transcriptional signature, differing in probability of firing and pulse duration, frequency, and intensity. In contrast to the prevailing view from both prokaryotes and eukaryotes that transcription displays binary behavior, strongly expressed housekeeping genes altered the magnitude of their transcriptional pulses during development. These nonbinary "tunable" responses may be better suited than stochastic switch behavior for housekeeping functions. Analysis of RNA synthesis kinetics using fluorescence recovery after photobleaching implied modulation of housekeeping-gene pulse strength occurs at the level of transcription initiation rather than elongation. In addition, disparities between single cell and population measures of transcript production suggested differences in RNA stability between gene classes. Analysis of stability using RNAseq revealed no major global differences in stability between developmental and housekeeping transcripts, although strongly induced RNAs showed unusually rapid decay, indicating tight regulation of expression.</p>
AB - <p>Transcription of genes can be discontinuous, occurring in pulses or bursts. It is not clear how properties of transcriptional pulses vary between different genes. We compared the pulsing of five housekeeping and five developmentally induced genes by direct imaging of single gene transcriptional events in individual living Dictyostelium cells. Each gene displayed its own transcriptional signature, differing in probability of firing and pulse duration, frequency, and intensity. In contrast to the prevailing view from both prokaryotes and eukaryotes that transcription displays binary behavior, strongly expressed housekeeping genes altered the magnitude of their transcriptional pulses during development. These nonbinary "tunable" responses may be better suited than stochastic switch behavior for housekeeping functions. Analysis of RNA synthesis kinetics using fluorescence recovery after photobleaching implied modulation of housekeeping-gene pulse strength occurs at the level of transcription initiation rather than elongation. In addition, disparities between single cell and population measures of transcript production suggested differences in RNA stability between gene classes. Analysis of stability using RNAseq revealed no major global differences in stability between developmental and housekeeping transcripts, although strongly induced RNAs showed unusually rapid decay, indicating tight regulation of expression.</p>
U2 - 10.1073/pnas.1117603109
DO - 10.1073/pnas.1117603109
M1 - Article
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 19
VL - 109
SP - 7350
EP - 7355
ER -