TY - JOUR
T1 - Chromosome and plasmid-borne P
LacO3O1
promoters differ in sensitivity to critically low temperatures
AU - Oliveira, Samuel M. D.
AU - Gonçalves, Nadia S. M.
AU - Kandavalli, Vinodh K.
AU - Martins, Leonardo
AU - Neeli-Venkata, Ramakanth
AU - Reyelt, Jan
AU - Fonseca, José M.
AU - Lloyd-Price, Jason
AU - Kranz, Harald
AU - Ribeiro, André S.
N1 - info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F88987%2F2012/PT#
UID/EEA/00066/203.
Academy of Finland (295027 to A.S.R. and 305342 to A.S.R.
Jane and Aatos Erkko Foundation (610536 to A.S.R.
PY - 2019/12/1
Y1 - 2019/12/1
N2 -
Temperature shifts trigger genome-wide changes in Escherichia coli’s gene expression. We studied if chromosome integration impacts on a gene’s sensitivity to these shifts, by comparing the single-RNA production kinetics of a P
LacO3O1
promoter, when chromosomally-integrated and when single-copy plasmid-borne. At suboptimal temperatures their induction range, fold change, and response to decreasing temperatures are similar. At critically low temperatures, the chromosome-integrated promoter becomes weaker and noisier. Dissection of its initiation kinetics reveals longer lasting states preceding open complex formation, suggesting enhanced supercoiling buildup. Measurements with Gyrase and Topoisomerase I inhibitors suggest hindrance to escape supercoiling buildup at low temperatures. Consistently, similar phenomena occur in energy-depleted cells by DNP at 30 °C. Transient, critically-low temperatures have no long-term consequences, as raising temperature quickly restores transcription rates. We conclude that the chromosomally-integrated P
LacO3O1
has higher sensitivity to low temperatures, due to longer-lasting super-coiled states. A lesser active, chromosome-integrated native lac is shown to be insensitive to Gyrase overexpression, even at critically low temperatures, indicating that the rate of escaping positive supercoiling buildup is temperature and transcription rate dependent. A genome-wide analysis supports this, since cold-shock genes exhibit atypical supercoiling-sensitivities. This phenomenon might partially explain the temperature-sensitivity of some transcriptional programs of E. coli.
AB -
Temperature shifts trigger genome-wide changes in Escherichia coli’s gene expression. We studied if chromosome integration impacts on a gene’s sensitivity to these shifts, by comparing the single-RNA production kinetics of a P
LacO3O1
promoter, when chromosomally-integrated and when single-copy plasmid-borne. At suboptimal temperatures their induction range, fold change, and response to decreasing temperatures are similar. At critically low temperatures, the chromosome-integrated promoter becomes weaker and noisier. Dissection of its initiation kinetics reveals longer lasting states preceding open complex formation, suggesting enhanced supercoiling buildup. Measurements with Gyrase and Topoisomerase I inhibitors suggest hindrance to escape supercoiling buildup at low temperatures. Consistently, similar phenomena occur in energy-depleted cells by DNP at 30 °C. Transient, critically-low temperatures have no long-term consequences, as raising temperature quickly restores transcription rates. We conclude that the chromosomally-integrated P
LacO3O1
has higher sensitivity to low temperatures, due to longer-lasting super-coiled states. A lesser active, chromosome-integrated native lac is shown to be insensitive to Gyrase overexpression, even at critically low temperatures, indicating that the rate of escaping positive supercoiling buildup is temperature and transcription rate dependent. A genome-wide analysis supports this, since cold-shock genes exhibit atypical supercoiling-sensitivities. This phenomenon might partially explain the temperature-sensitivity of some transcriptional programs of E. coli.
UR - http://www.scopus.com/inward/record.url?scp=85062972167&partnerID=8YFLogxK
U2 - 10.1038/s41598-019-39618-z
DO - 10.1038/s41598-019-39618-z
M3 - Article
C2 - 30872616
AN - SCOPUS:85062972167
SN - 2045-2322
VL - 9
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 4486
ER -