Darlene Cabanas, Jan Zarzycki-Siek, Andrew P Bluhm, Zhenxin Sun, Michael J Schurr, Yun Heacock-Kang, Christopher D Sibley, Dawson Fogen, Michael H Norris, Ian A McMillan, Hung Vo, Herbert P Schweizer, Shawn Lewenza, Bradley R Borlee, Tung T Hoang, Stuart P Donachie
Bacterial cooperative associations and dynamics in the biofilm microenvironments are of special interest in recent years. Knowledge of localized gene-expression and corresponding bacterial behaviors within the biofilm architecture at a global scale has been limited, due to a lack of robust technology to study limited number of cells in stratified layers of biofilms. With our recent pioneering developments in single bacterial cell transcriptomic analysis technology, we generated herein an unprecedented spatial transcriptome map of the mature in vitro Pseudomonas aeruginosa biofilm model, revealing contemporaneous yet altered bacterial behaviors at different layers within the biofilm architecture (i.e., surface, middle, and interior of the biofilm). Many genes encoding unknown functions were highly expressed at the biofilm-solid interphase, exposing a critical gap in the knowledge of their activities that maybe unique to this interior niche. Several genes of unknown functions are critical for biofilm formation. The in vivo importance of these unknown proteins was validated in invertebrate (fruit fly) and vertebrate (mouse) models. We envisage the future value of this report to the community, in aiding the further pathophysiological understanding of P. aeruginosa biofilms. Our approach will open doors to the study of bacterial functional genomics of different species in numerous settings. This article is protected by copyright. All rights reserved.
A comprehensive spatial transcriptome map of the mature Pseudomonas aeruginosa biofilm was generated, revealing contemporaneous yet altered bacterial behaviors at different layers within the biofilm architecture. Many genes encoding unknown proteins were highly expressed at the interior of biofilm. Several genes of unknown functions are critical for biofilm formation and in vivo fitness in fruit fly and mouse infection models. We envisage this report aiding the further pathophysiological understanding of P. aeruginosa biofilms.