PT  - JOURNAL ARTICLE
AU  - Tufail Bashir
AU  - Rowan D Brackston
AU  - Christopher J Waite
AU  - Ioly Kotta-Loizou
AU  - Christoph Engl
AU  - Martin Buck
AU  - Jörg Schumacher
TI  - Molecular origins of transcriptional heterogeneity in diazotrophic <em>K. oxytoca</em>
AID  - 10.1101/2020.02.18.955476
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.02.18.955476
4099  - http://biorxiv.org/content/early/2020/02/20/2020.02.18.955476.short
4100  - http://biorxiv.org/content/early/2020/02/20/2020.02.18.955476.full
AB  - Phenotypic heterogeneity in clonal bacterial batch cultures is an important adaptive strategy to changing environments, including in diazotrophs with the unique capacity to convert di-nitrogen into bio-available ammonium. In diazotrophic Klebsiella oxytoca we simultaneously measured mRNA levels of key regulatory (glnK-amtB, nifLA) and structural (nifHDK) operons required for establishing nitrogen fixation, using dual molecule, single cell RNA-FISH. Through stochastic transcription models and mutual information analysis we revealed likely molecular origins for heterogeneity in nitrogenase expression. In wildtype and regulatory variant strains we inferred contributions from intrinsic and extrinsic noise, finding that nifHDK transcription is inherently bursty, but that noise propagation through signalling is also significant. The regulatory gene glnK had the highest discernible effect on nifHDK variance, while noise from factors outside of the regulatory pathway were negligible. Results provide evidence that heterogeneity is a fundamental property of this regulatory system, indicating potential constraints for engineering homogeneous nitrogenase expression.