Regulatory Connections Between the Cyanobacterial Factor PipX and the Ribosome Assembly GTPase EngA

Cyanobacteria, phototrophic organisms performing oxygenic photosynthesis, must adapt their metabolic processes to important environmental challenges, like those imposed by the succession of days and nights. Not surprisingly, certain regulatory proteins are found exclusively in this phylum. One of these unique proteins, PipX, provides a mechanistic link between signals of carbon/nitrogen and of energy, transduced by the signaling protein PII, and the control of gene expression by the global nitrogen regulator NtcA. PII, required for cell survival unless PipX is inactivated or downregulated, functions by protein–protein interactions with transcriptional regulators, transporters, and enzymes. PipX also functions by protein–protein interactions, and previous studies suggested the existence of additional interacting partners or included it into a relatively robust six-node synteny network with proteins apparently unrelated to the nitrogen regulation system. To investigate additional functions of PipX while providing a proof of concept for the recently developed cyanobacterial linkage network, here we analyzed the physical and regulatory interactions between PipX and an intriguing component of the PipX synteny network, the essential ribosome assembly GTPase EngA. The results provide additional insights into the functions of cyanobacterial EngA and of PipX, showing that PipX interacts with the GD1 domain of EngA in a guanosine diphosphate-dependent manner and interferes with EngA functions in Synechococcus elongatus at a low temperature, an environmentally relevant context. Therefore, this work expands the PipX interaction network and establishes a possible connection between nitrogen regulation and the translation machinery. We discuss a regulatory model integrating previous information on PII–PipX with the results presented in this work.

Glutamine involvement in nitrogen regulation of cellulase production in fungi

Background Cellulase synthesized by fungi can environment-friendly and sustainably degrades cellulose to fermentable sugars for producing cellulosic biofuels, biobased medicine and fine chemicals. Great efforts have been made to study the regulation mechanism of cellulase biosynthesis in fungi with the focus on the carbon sources, while little attention has been paid to the impact and regulation mechanism of nitrogen sources on cellulase production. Results Glutamine displayed the strongest inhibition effect on cellulase biosynthesis in Trichoderma reesei, followed by yeast extract, urea, tryptone, ammonium sulfate and l-glutamate. Cellulase production, cell growth and sporulation in T. reesei RUT-C30 grown on cellulose were all inhibited with the addition of glutamine (a preferred nitrogen source) with no change for mycelium morphology. This inhibition effect was attributed to both l-glutamine itself and the nitrogen excess induced by its presence. In agreement with the reduced cellulase production, the mRNA levels of 44 genes related to the cellulase production were decreased severely in the presence of glutamine. The transcriptional levels of genes involved in other nitrogen transport, ribosomal biogenesis and glutamine biosynthesis were decreased notably by glutamine, while the expression of genes relevant to glutamate biosynthesis, amino acid catabolism, and glutamine catabolism were increased noticeably. Moreover, the transcriptional level of cellulose signaling related proteins ooc1 and ooc2, and the cellular receptor of rapamycin trFKBP12 was increased remarkably, whose deletion exacerbated the cellulase depression influence of glutamine. Conclusion Glutamine may well be the metabolite effector in nitrogen repression of cellulase synthesis, like the role of glucose plays in carbon catabolite repression. Glutamine under excess nitrogen condition repressed cellulase biosynthesis significantly as well as cell growth and sporulation in T. reesei RUT-C30. More importantly, the presence of glutamine notably impacted the transport and metabolism of nitrogen. Genes ooc1, ooc2, and trFKBP12 are associated with the cellulase repression impact of glutamine. These findings advance our understanding of nitrogen regulation of cellulase production in filamentous fungi, which would aid in the rational design of strains and fermentation strategies for cellulase production in industry.

Evolutionary loss of thermal acclimation accompanied by periodic monocarpic mass flowering in Strobilanthes flexicaulis

While life history, physiology and molecular phylogeny in plants have been widely studied, understanding how physiology changes with the evolution of life history change remains largely unknown. In two closely related understory Strobilanthes plants, the molecular phylogeny has previously shown that the monocarpic 6-year masting S. flexicaulis have evolved from a polycarpic perennial, represented by the basal clade S. tashiroi. The polycarpic S. tashiroi exhibited seasonal thermal acclimation with increased leaf respiratory and photosynthetic metabolism in winter, whereas the monocarpic S. flexicaulis showed no thermal acclimation. The monocarpic S. flexicaulis required rapid height growth after germination under high intraspecific competition, and the respiration and N allocation were biased toward nonphotosynthetic tissues. By contrast, in the long-lived polycarpic S. tashiroi, these allocations were biased toward photosynthetic tissues. The life-history differences between the monocarpic S. flexicaulis and the polycarpic S. tashiroi are represented by the “height growth” and “assimilation” paradigms, respectively, which are controlled by different patterns of respiration and nitrogen regulation in leaves. The obtained data indicate that the monocarpic S. flexicaulis with the evolutionary loss of thermal acclimation may exhibit increased vulnerability to global warming.

Evolutionary loss of thermal acclimation accompanied by periodic monocarpic mass flowering in Strobilanthes species

While life history, physiology and molecular phylogeny in plants have been widely studied, understanding how physiology changes with the evolutionally life historical change remains largely unknown. In two closely related understory Strobilanthes plants, the molecular phylogeny has previously shown that the monocarpic 6-year masting S. flexicaulis have evolved from a polycarpic perennial, represented by the basal clade S. tashiroi. The polycarpic S. tashiroi exhibited seasonal thermal acclimation with increased leaf respiratory and photosynthetic metabolism in winter, whereas the monocarpic S. flexicaulis showed no thermal acclimation. The monocarpic S. flexicaulis required rapid height growth after germination under high intraspecific competition, and the respiration and N allocation were biased toward nonphotosynthetic tissues. By contrast, in the long-lived polycarpic S. tashiroi, these allocations were biased toward photosynthetic tissues. The life-history differences between the monocarpic S. flexicaulis and the polycarpic S. tashiroi are represented by the “height growth” and “assimilation” paradigms, respectively, which are controlled by different patterns of respiration and nitrogen regulation in leaves. The obtained data suggest that the monocarpic S. flexicaulis with the evolutionary loss of thermal acclimation will more often exhibit increased vulnerability to global warming.