Development of a systems-level model of cellular dynamics in response to different environmental conditions in cyanobacteria
EMSL Project ID
44624
Abstract
This proposal will utilize a systems biology approach to develop a knowledgebase of the metabolic and regulatory networks that can ultimately be used for the photobiological production of advanced biofuels and/or their chemical precursors by cyanobacteria. We will focus on two cyanobacterial species: Synechocystis sp. PCC 6803 and Cyanothece sp. ATCC 51142. Synechocystis 6803 is a fresh water organism that is arguably the most widely studied cyanobacterium at the current time. The first photosynthetic organism with a completely sequenced genome, Synechocystis is naturally transformable, so that targeted gene replacements can be readily generated. Cyanothece is a marine cyanobacterium that can perform oxygenic photosynthesis and, unlike Synechocystis, can also fix nitrogen. Despite this major metabolic difference, the two strains share many similarities at the genomic and proteomic levels. This complementarity has permitted us to use the two strains as partners in our experiments, and we have extensive experience in studying both organisms. In the present proposal, our objective is to characterize the cellular response network that govern bioenergy production and cell morphology of cyanobacteria under different environmental conditions. We propose two interconnected sets of experiments to gain this understanding. The use of sophisticated instrumentation available at EMSL will be vital in achieving these goals. Both PI's labs have extensive experience collaborating with key investigators at EMSL/PNNL over the past 5 years, resulting in numerous publications. Part 1. Examining the effect of environmental conditions on cellular morphology in cyanobacteria Objective: We aim to study the organization of protein complexes within the photosynthetic and cytoplasmic membranes, and the relationship of the membranes to the many metabolic storage granules found throughout the cell. This project will enable us to determine, with great precision, the amount and intracellular location of many chemical components. Cell sorting will allow us to enrich for cell populations displaying desired characteristics and thus more uniform populations for examination.
Part 2. Gaining a systems level understanding of the physiological changes under different nutritional conditions by combining transcriptomic, metabolomic, metalomic, and proteomic datasets Objective: We aim to develop a system level understanding of the physiological changes under conditions that mimic natural environments as well as to maximize biomass and biofuel production. Significance: This project represents an unparalleled and unique opportunity to understand the structure and function of an important biofuel-producing organism at both the sub-cellular and cellular levels. With this information, we will be able to construct a predictive model of the cell and its metabolic fluxes and thus to ask key questions for future experiments. The use of the EMSL capabilities is essential for this work, because the staff has developed the scientific expertise to interpret the experimental subtleties. In turn, the successful conclusion of this project will demonstrate the importance of having this collection of state-of-the-art capabilities at EMSL, while also answering one key scientific challenge relevant to the DOE. The lab will be responsible for dissecting the metabolic, transcriptional and structural organization of an organism that harnesses solar energy and is capable of producing many alternative forms of energy.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2011-10-01
End Date
2014-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Aryal UK, SJ Callister, BH McMahon, LA McCue, JN Brown, J Stockel, ML Liberton, S Mishra, X Zhang, CD Nicora, TE Angel, DW Koppenaal, RD Smith, HB Pakrasi, and LA Sherman. 2014. "Proteomic profiles of five strains of oxygenic photosynthetic cyanobacteria of the genus Cyanothece." Journal of Proteome Research 13(7):3262–3276. doi:10.1021/pr5000889
Aryal UK, SJ Callister, S Mishra, X Zhang, JI Shutthanandan, AK Shukla, ME Monroe, RJ Moore, DW Koppenaal, RD Smith, and L Sherman. 2013. "Proteome Analyses of Strains ATCC 51142 and PCC 7822 of the Diazotrophic Cyanobacterium Cyanothece sp under Culture Conditions Resulting in Enhanced H-2 Production." Applied and Environmental Microbiology 79(4):1070-1077. doi:10.1128/AEM.02864-12. Epub 2012 Nov 30.
Welkie DG, DM Sherman, WB Chrisler, G Orr, and LA Sherman. 2013. "Analysis of carbohydrate storage granules in the diazotrophic cyanobacterium Cyanothece sp. PCC 7822." Photosynthesis Research 118(1-2):25-36. doi:10.1007/s11120-013-9941-z
Welkie DG, X Zhang, LM Markillie, RC Taylor, G Orr, JM Jacobs, K Bhide, J Thimmapuram, MA Gritsenko, HD Mitchell, RD Smith, and LA Sherman. 2014. "Transcriptomic and proteomic dynamics in the metabolism of a diazotrophic cyanobacterium, Cyanothece sp. PCC 7822 during a diurnal light-dark cycle." BMC Genomics 15:1185. doi:10.1186/1471-2164-15-1185