Dynamic competition between a ligand and transcription factor NusA governs riboswitch-mediated transcription regulation
Chauvier, Adrien; Ajmera, Pujan; Yadav, Rajeev; Walter, Nils
2021-10-14
View/ Open
EC25-No Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at EC-25 in the absence of Fluoride
(3.1MB
application/zip)NusA binding assay - Single molecule traces obtained at EC-25 in the absence of Fluoride
EC25-With Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at EC-25 in the presence of Fluoride
(503.6KB
application/zip)NusA binding assay - Single molecule traces obtained at EC-25 in the presence of Fluoride
HisP - NusA.zip
NusA binding assay - Single molecule traces obtained at HisP
(425.6KB
application/zip)NusA binding assay - Single molecule traces obtained at HisP
PEC48-A10U-No Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-A10U mutant in the absence of Fluoride
(500.5KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-A10U mutant in the absence of Fluoride
PEC48-A10U-U38A-No Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-A10U-U38A mutant in the absence of Fluoride
(347.1KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-A10U-U38A mutant in the absence of Fluoride
PEC48-A10U-U38A-With Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-A10U-U38A mutant in the presence of Fluoride
(155.3KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-A10U-U38A mutant in the presence of Fluoride
PEC48-A10U-With Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-A10U mutant in the presence of Fluoride
(360.4KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-A10U mutant in the presence of Fluoride
PEC48-U38A-No Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-U38A mutant in the absence of Fluoride
(504.9KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-U38A mutant in the absence of Fluoride
PEC48-U38A-With Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-U38A mutant in the presence of Fluoride
(636.7KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-U38A mutant in the presence of Fluoride
PEC48-WT-No Fluoride - Competition unlabeled NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-WT in the presence of unlabeled NusA
(413.6KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-WT in the presence of unlabeled NusA
PEC48-WT-No Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-WT in the absence of Fluoride
(751.7KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-WT in the absence of Fluoride
PEC48-WT-With Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC48-WT in the presence of Fluoride
(827.9KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC48-WT in the presence of Fluoride
PEC72-No Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC72 in the absence of Fluoride
(238.4KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC72 in the absence of Fluoride
PEC72-With Fluoride - NusA.zip
NusA binding assay - Single molecule traces obtained at PEC72 in the presence of Fluoride
(405.2KB
application/zip)NusA binding assay - Single molecule traces obtained at PEC72 in the presence of Fluoride
PEC72-No Fluoride - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at PEC72 in the absence of Fluoride
(325.9KB
application/zip)SiM-KARTS - Single molecule traces obtained at PEC72 in the absence of Fluoride
PEC72-With Fluoride - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at PEC72 in the presence of Fluoride
(255KB
application/zip)SiM-KARTS - Single molecule traces obtained at PEC72 in the presence of Fluoride
RealTime Trx-No Fluoride-25micros rNTPs - 100nM NusA.zip
Real Time transcription - 25µM rNTPs with 100nM NusA
(106KB
application/zip)Real Time transcription - 25µM rNTPs with 100nM NusA
RealTime Trx-No Fluoride-25micros rNTPs - No NusA.zip
Real Time transcription - 25µM rNTPs
(152.3KB
application/zip)Real Time transcription - 25µM rNTPs
RealTime Trx-No Fluoride-100micros rNTPs - 100nM NusA.zip
Real Time transcription - 100µM rNTPs with 100nM NusA
(124.9KB
application/zip)Real Time transcription - 100µM rNTPs with 100nM NusA
RealTime Trx-No Fluoride-100micros rNTPs - No NusA.zip
Real Time transcription - 100µM rNTPs
(139.6KB
application/zip)Real Time transcription - 100µM rNTPs
RealTime Trx-No Fluoride-250micros rNTPs - 100nM NusA.zip
Real Time transcription - 250µM rNTPs with 100nM NusA
(150.7KB
application/zip)Real Time transcription - 250µM rNTPs with 100nM NusA
RealTime Trx-No Fluoride-250micros rNTPs - No NusA.zip
Real Time transcription - 250µM rNTPs
(79.7KB
application/zip)Real Time transcription - 250µM rNTPs
RealTime Trx-U38A-No Fluoride-25micros rNTPs - 2nM NusA-Cy5.zip
Real Time transcription - U38A mutant - 25µM rNTPs with 2nM NusA-Cy5 in the absence of Fluoride
(402.8KB
application/zip)Real Time transcription - U38A mutant - 25µM rNTPs with 2nM NusA-Cy5 in the absence of Fluoride
RealTime Trx-U38A-With Fluoride-25micros rNTPs - 2nM NusA-Cy5.zip
Real Time transcription - U38A mutant - 25µM rNTPs with 2nM NusA-Cy5 in the presence of Fluoride
(223.6KB
application/zip)Real Time transcription - U38A mutant - 25µM rNTPs with 2nM NusA-Cy5 in the presence of Fluoride
RealTime Trx-With Fluoride-25micros rNTPs - No NusA.zip
Real Time transcription - 25µM rNTPs in the presence of Fluoride
(126.9KB
application/zip)Real Time transcription - 25µM rNTPs in the presence of Fluoride
RealTime Trx-WT-No Fluoride-25micros rNTPs - 2nM NusA-Cy5.zip
Real Time transcription - WT - 25µM rNTPs with 2nM NusA-Cy5 in the absence of Fluoride
(240.3KB
application/zip)Real Time transcription - WT - 25µM rNTPs with 2nM NusA-Cy5 in the absence of Fluoride
RealTime Trx-WT-With Fluoride-25micros rNTPs - 2nM NusA-Cy5.zip
Real Time transcription - WT - 25µM rNTPs with 2nM NusA-Cy5 in the presence of Fluoride
(256.4KB
application/zip)Real Time transcription - WT - 25µM rNTPs with 2nM NusA-Cy5 in the presence of Fluoride
RT72-No Fluoride-1mM rNTPs - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the absence of Fluoride - 1mM rNTPs
(285.4KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the absence of Fluoride - 1mM rNTPs
RT72-No Fluoride-1mM rNTPs-With NusA - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the absence of Fluoride - 1mM rNTPs with NusA
(186.3KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the absence of Fluoride - 1mM rNTPs with NusA
RT72-No Fluoride-100micros rNTPs - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the absence of Fluoride - 100µM rNTPs
(434.2KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the absence of Fluoride - 100µM rNTPs
RT72-With Fluoride CoTrx-1mM rNTPs - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride (Co-Trx) - 1mM rNTPs
(104.8KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride (Co-Trx) - 1mM rNTPs
RT72-With Fluoride PostTrx-1mM rNTPs - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride (Post-Trx) - 1mM rNTPs
(243.1KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride (Post-Trx) - 1mM rNTPs
RT72-With Fluoride-1mM rNTPs-With NusA - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride - 1mM rNTPs with NusA
(167.5KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride - 1mM rNTPs with NusA
RT72-With Fluoride-100micros rNTPs - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride - 100µM rNTPs
(727.8KB
application/zip)SiM-KARTS - Single molecule traces obtained at RT72 in the presence of Fluoride - 100µM rNTPs
Terminator Only-1mM rNTPs - SiMKARTS.zip
SiM-KARTS - Single molecule traces obtained at the terminator only construct - 1mM rNTPs
(324.4KB
application/zip)SiM-KARTS - Single molecule traces obtained at the terminator only construct - 1mM rNTPs
Abstract
Co-transcriptional RNA folding is widely assumed to influence the timely control of gene expression, but our understanding remains limited. In bacteria, the fluoride (F-) sensing riboswitch is a transcriptional control element essential to defend against toxic F- levels. Using this model riboswitch, we find that its ligand F- and essential bacterial transcription factor NusA compete for binding the co-transcriptionally folding RNA, opposing each other’s modulation of downstream pausing and termination by RNA polymerase. Single molecule fluorescence assays probing active transcription elongation complexes discover that NusA unexpectedly binds highly reversibly, frequently interrogating the complex for emerging, co-transcriptionally folding RNA duplexes. NusA thus fine-tunes the transcription rate in dependence of the ligand-responsive higher order structure of the riboswitch. At the high NusA concentrations found intracellularly, this dynamic modulation is expected to lead to adaptive bacterial transcription regulation with fast response times.Deep Blue DOI
Subjects
Co-transcriptional folding, RNA polymerase pausing, NusA, riboswitch, single molecule fluorescence microscopy
Types
Article
Metadata
Show full item recordRemediation of Harmful Language
The University of Michigan Library aims to describe library materials in a way that respects the people and communities who create, use, and are represented in our collections. Report harmful or offensive language in catalog records, finding aids, or elsewhere in our collections anonymously through our metadata feedback form. More information at Remediation of Harmful Language.
Accessibility
If you are unable to use this file in its current format, please select the Contact Us link and we can modify it to make it more accessible to you.