We have developed a theory that lets us calculate the efficiency of microbes that absorb electricity and store CO 2 as biofuels with greater efficiency than photosynthesis. We outline 10 development scenarios including re-engineering direct electron uptake microbes with high-efficiency CO 2 fixation; scaling up H 2oxidizing microbe systems to store megawatts of electricity; engineering direct electron uptake microbes to make highly conductive artificial biofilms to enable high power density electricity storage; and engineering microbes that assimilate electrochemically reduced CO 2 with electron uptake.
Here we describe the protein interaction platform assay, a method for identifying interacting proteins in Saccharomyces cerevisiae. This assay relies on the reovirus scaffolding protein μNS, which forms large focal inclusions in living cells. When a query protein is fused to μNS and potential interaction partners are fused to a fluorescent reporter, interactors can be identified by screening for yeast that display fluorescent foci.A variety of methods have been developed to screen for interacting proteins in living cells, including yeast two-hybrid (Y2H) and protein-fragment complementation assays1 , 2. We recently demon-strated a complementary approach: the protein interaction plat-form (PIP) assay, in which we generate inclusions that serve as platforms for assembling interacting proteins in living mammalian cells. Here, we investigated whether this assay could be adapted to the yeast S. cerevisiae, in which (unlike in mammalian systems) plasmids for expressing proteins are readily maintained.PIP relies on the reovirus scaffolding protein μNS to form distinctively large focal cytoplasmic inclusions in mammalian cells3 ,4 . The C-terminal one-third of μNS (μNS residues 471-721) formed inclusions when expressed in S. cerevisiae (Fig. 1a). These inclusions did not perturb yeast growth, and mutations in the putative zinc-hook motif of μNS that disrupt the formation of platforms in mammalian cells 5 also disrupted their formation in yeast (Fig. 1b).The basic design of PIP as adapted to yeast involved fusing one protein (A) to μNS and a second protein (B) to GFP, so that if A and B interact, fluorescent green inclusions are visible in individual yeast cells (Fig. 1c,d). We created a series of recombination-based destination vectors using Gateway technology (Invitrogen) to facilitate the transfer of open reading frames such that they are fused to μNS at their N termini and expressed from either a constitutive (GPD) or an inducible (GAL1) yeast promoter. Many Gram-negative bacterial pathogens, including Shigella spp. and Salmonella spp., directly inject virulence proteins, called effectors, into host cells via specialized secretion systems. Some effectors require chaperones for delivery to the bacterial secretion apparatus. Shigella flexneri encodes four chaperones and at least 30
Under nutrient-limiting conditions, plants will enter into symbiosis with arbuscular mycorrhizal (AM) fungi for the enhancement of mineral nutrient acquisition from the surrounding soil. AM fungi live in close, intracellular association with plant roots where they transfer phosphate and nitrogen to the plant in exchange for carbon. They are obligate fungi, relying on their host as their only carbon source. Much has been discovered in the last decade concerning the signaling events during initiation of the AM symbiosis, including the identification of signaling molecules generated by both partners. This signaling occurs through symbiosis-specific gene products in the host plant, which are indispensable for normal AM development. At the same time, plants have adapted complex mechanisms for avoiding infection by pathogenic fungi, including an innate immune response to general microbial molecules, such as chitin present in fungal cell walls. How it is that AM fungal colonization is maintained without eliciting a defensive response from the host is still uncertain. In this review, we present a summary of the molecular signals and their elicited responses during initiation of the AM symbiosis, including plant immune responses and their suppression.Keywords: Arbuscular mycorrhizal fungi; chitin; chitooligosaccharide; cutin; LysM; MAMP-triggered immunity; Myc-LCO; Nod factor; strigolactones; symbiosis Citation: Schmitz AM, Harrison MJ (2014) Signaling events during initiation of arbuscular mycorrhizal symbiosis.
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