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Series GSE234096 Query DataSets for GSE234096
Status Public on Jun 07, 2023
Title The Caulobacter NtrB-NtrC two-component system bridges nitrogen assimilation and cell development [ChIP-seq]
Organism Caulobacter vibrioides NA1000
Experiment type Genome binding/occupancy profiling by high throughput sequencing
Summary Caulobacter species are dimorphic Gram-negative bacteria that inhabit diverse terrestrial and aquatic ecosystems. A suite of molecular sensory systems enables these microbes to control growth, development, and reproduction in response to levels of essential elements, including carbon, nitrogen, and phosphorus. Although the enhancer binding protein NtrC and its cognate sensor histidine kinase NtrB are well-established regulators of bacterial nitrogen assimilation, their precise functions in Caulobacter metabolism and cell development remained largely undefined. Deletion of C. crescentus ntrC slowed cell growth in complex medium, while ntrB and ntrC were essential for growth when ammonium was the sole nitrogen source due to their requirement for glutamine synthase (glnA) expression. Random transposition of a conserved IS3-family mobile genetic element frequently rescued the growth defect of ntrC mutant strains by restoring glnBA transcription, revealing a possible role for IS3 transposition in shaping the evolution of Caulobacter populations during nitrogen limitation. We further defined the NtrC regulon through transcriptomic and ChIP-seq analyses, which demonstrated the role of NtrC as both a transcriptional activator and repressor. The chromosome of C. crescentus harbors dozens of direct binding sites for NtrC, with a significant proportion located near genes involved in polysaccharide biosynthesis. Numerous NtrC binding sites align with those of GapR, an essential protein involved in chromosome organization, and MucR1, a cell cycle regulator. This observation suggests that NtrC participates in the regulation of cell cycle and cell development. Indeed, loss of NtrC function led to elongated polar stalks and elevated synthesis of cell envelope polysaccharides. These developmental defects were restored by supplementing media with glutamine or by ectopic expression of the glnBA operon. This study establishes a regulatory connection between NtrC, nitrogen metabolism, polar morphogenesis, and envelope polysaccharide synthesis in Caulobacter.
Overall design To investigate the NtrC direct regulon/DNA binding sites in Caulobacter by creating a 3xFLAG-tagged ntrC allele, which resulted in a 3xFLAG tag at the N-terminus of NtrC
We performed NtrC DNA-binding analysis from ChIP-seq data of Caulobacter harboring 3xFLAG-ntrC grown in PYE to log phase
Contributor(s) North H, McLaughlin M, Fiebig A, Crosson S
Citation(s) 37791753
Submission date Jun 05, 2023
Last update date Nov 09, 2023
Contact name Sean Crosson
Phone 5178845345
Organization name Michigan State University
Department Dept. Microbiology and Molecular Genetics
Street address 567 Wilson Rd
City East Lansing
State/province Michigan
ZIP/Postal code 48824
Country USA
Platforms (1)
GPL33463 NextSeq 2000 (Caulobacter vibrioides NA1000)
Samples (2)
GSM7445764 ∆ntrC xylX::pPTM057-3xFLAG-ntrC input DNA
GSM7445765 ∆ntrC xylX::pPTM057-3xFLAG-ntrC output DNA
This SubSeries is part of SuperSeries:
GSE234097 The Caulobacter NtrB-NtrC two-component system bridges nitrogen assimilation and cell development
BioProject PRJNA980111

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Supplementary file Size Download File type/resource
GSE234096_Galaxy11-_SCD346_345_.bigwig 6.1 Mb (ftp)(http) BIGWIG
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Raw data are available in SRA
Processed data are available on Series record

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