title: Transcriptomic and proteomic dynamics in the metabolism of a diazotrophic cyanobacterium, Cyanothece sp. PCC 7822 during a diurnal light-dark cycle.
abstract: BACKGROUND
  Cyanothece sp. PCC 7822 is an excellent cyanobacterial model organism with great potential to be applied as a biocatalyst for the production of high value compounds. Like other unicellular diazotrophic cyanobacterial species, it has a tightly regulated metabolism synchronized to the light-dark cycle. Utilizing transcriptomic and proteomic methods, we quantified the relationships between transcription and translation underlying central and secondary metabolism in response to nitrogen free, 12 hour light and 12 hour dark conditions.
  
  
  RESULTS
  By combining mass-spectrometry based proteomics and RNA-sequencing transcriptomics, we quantitatively measured a total of 6766 mRNAs and 1322 proteins at four time points across a 24 hour light-dark cycle. Photosynthesis, nitrogen fixation, and carbon storage relevant genes were expressed during the preceding light or dark period, concurrent with measured nitrogenase activity in the late light period. We describe many instances of disparity in peak mRNA and protein abundances, and strong correlation of light dependent expression of both antisense and CRISPR-related gene expression. The proteins for nitrogenase and the pentose phosphate pathway were highest in the dark, whereas those for glycolysis and the TCA cycle were more prominent in the light. Interestingly, one copy of the psbA gene encoding the photosystem II (PSII) reaction center protein D1 (psbA4) was highly upregulated only in the dark. This protein likely cannot catalyze O2 evolution and so may be used by the cell to keep PSII intact during N2 fixation. The CRISPR elements were found exclusively at the ends of the large plasmid and we speculate that their presence is crucial to the maintenance of this plasmid.
  
  
  CONCLUSIONS
  This investigation of parallel transcriptional and translational activity within Cyanothece sp. PCC 7822 provided quantitative information on expression levels of metabolic pathways relevant to engineering efforts. The identification of expression patterns for both mRNA and protein affords a basis for improving biofuel production in this strain and for further genetic manipulations. Expression analysis of the genes encoded on the 6 plasmids provided insight into the possible acquisition and maintenance of some of these extra-chromosomal elements.
authors.forename: David
authors.initials: D
authors.lastname: Welkie
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zhang
authors.forename: Meng Lye
authors.initials: ML
authors.lastname: Markillie
authors.forename: Ronald
authors.initials: R
authors.lastname: Taylor
authors.forename: Galya
authors.initials: G
authors.lastname: Orr
authors.forename: Jon
authors.initials: J
authors.lastname: Jacobs
authors.forename: Ketaki
authors.initials: K
authors.lastname: Bhide
authors.forename: Jyothi
authors.initials: J
authors.lastname: Thimmapuram
authors.forename: Marina
authors.initials: M
authors.lastname: Gritsenko
authors.forename: Hugh
authors.initials: H
authors.lastname: Mitchell
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
authors.affiliation: Department of Biological Sciences, Purdue University, West Lafayette, IN, USA. sherman@purdue.edu.
authors.forename: Louis A
authors.initials: LA
authors.lastname: Sherman
mesh_terms: D056804:Biofuels / Q000382:microbiology; D002940:Circadian Rhythm / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D046889:Cyanothece / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology / Q000528:radiation effects; D003624:Darkness; D020869:Gene Expression Profiling; D009586:Nitrogen Fixation / Q000235:genetics / Q000528:radiation effects; D010788:Photosynthesis / Q000235:genetics / Q000528:radiation effects; D014176:Protein Biosynthesis / Q000528:radiation effects; D040901:Proteomics; D016372:RNA, Antisense / Q000235:genetics; D014158:Transcription, Genetic / Q000528:radiation effects
chemical_list: D056804:Biofuels; D016372:RNA, Antisense