Project description:Change in gene expression for a wild-type (Nostoc punctiforme ATCC 29133) and hmpD-deletion strain (UCD 543) of Nostoc punctiforme ATCC 29133 over the time course of hormogonium development This study is further descirbed in Risser, D.D. and Meeks, J.C. 2013. Comparative transcriptomics with a motility deficient mutant leads to identification of a novel polysaccharide secretion system in Nostoc punctiforme. Molecular Microbiology
Project description:Change in gene expression for a wild-type (Nostoc punctiforme ATCC 29133) and hmpD-deletion strain (UCD 543) of Nostoc punctiforme ATCC 29133 over the time course of hormogonium development This study is further descirbed in Risser, D.D. and Meeks, J.C. 2013. Comparative transcriptomics with a motility deficient mutant leads to identification of a novel polysaccharide secretion system in Nostoc punctiforme. Molecular Microbiology Total RNA from 3 biological replicates at each time point from 0 to 24 hours after hormogonium induction was converted to cDNA, dye-labled and hybridized to nimblegen 12x135k array slides
Project description:Heterocysts, cells specialized for nitrogen fixation in certain filamentous cyanobacteria, appear singly in a nonrandom spacing pattern along the chain of vegetative cells. A two-stage, biased initiation and competitive resolution model has been proposed to explain the establishment of this spacing pattern. There is substantial evidence that competitive resolution of a subset of cells initiating differentiation occurs by interactions between a self enhancing activator, HetR, and a diffusible inhibitor PatS-5 (RGSGR). Results presented here show that the absence of a unique membrane protein, PatN, in Nostoc punctiforme strain ATCC 29133 leads to a threefold increase in heterocyst frequency and a fourfold decrease in the vegetative cell interval between heterocysts. A PatN-GFP translational fusion shows a pattern of biased inheritance in daughter vegetative cells of ammonium-grown cultures. Inactivation of another heterocyst patterning gene, patA, is epistatic to inactivation of patN, and transcription of patA increases in a patN- deletion strain, implying that patN may function by modulating levels of patA. The presence of PatN is hypothesized to decrease the competency of a vegetative cell to initiate heterocyst differentiation, and the cellular concentration of PatN is dependent on cell division that results in cells transiently depleted of PatN. We suggest that biased inheritance of cell-fate determinants is a phylogenetic domain- spanning paradigm in the development of biological patterns. Change in gene expression for a wild-type (UCD 153) and patN-deletion strain, (UCD 524) of Nostoc punctiforme ATCC 29133 over the time course of heterocyst developments. Total RNA from 3 biological replicates at each time point from 0 to 120 hours after removal of combined nitrogen (Nitrogen step-down) was converted to cDNA, dye-labled and hybridized to nimblegen 12x135k array slides.
Project description:Cyanobacteria are a rich source of natural products and are known to produce terpenoids. These bacteria are the major source of the musty-smelling terpenes geosmin and 2-methylisoborneol, which are found in many natural water supplies; however, no terpene synthases have been characterized from these organisms to date. Here, we describe the characterization of three sesquiterpene synthases identified in Nostoc sp. strain PCC 7120 (terpene synthase NS1) and Nostoc punctiforme PCC 73102 (terpene synthases NP1 and NP2). The second terpene synthase in N. punctiforme (NP2) is homologous to fusion-type sesquiterpene synthases from Streptomyces spp. shown to produce geosmin via an intermediate germacradienol. The enzymes were functionally expressed in Escherichia coli, and their terpene products were structurally identified as germacrene A (from NS1), the eudesmadiene 8a-epi-alpha-selinene (from NP1), and germacradienol (from NP2). The product of NP1, 8a-epi-alpha-selinene, so far has been isolated only from termites, in which it functions as a defense compound. Terpene synthases NP1 and NS1 are part of an apparent minicluster that includes a P450 and a putative hybrid two-component protein located downstream of the terpene synthases. Coexpression of P450 genes with their adjacent located terpene synthase genes in E. coli demonstrates that the P450 from Nostoc sp. can be functionally expressed in E. coli when coexpressed with a ferredoxin gene and a ferredoxin reductase gene from Nostoc and that the enzyme oxygenates the NS1 terpene product germacrene A. This represents to the best of our knowledge the first example of functional expression of a cyanobacterial P450 in E. coli.
