The Role of Chitinase Production by Stenotrophomonas maltophilia Strain C3 in Biological Control of Bipolaris sorokiniana

The role of chitinase production by Stenotrophomonas maltophilia strain C3 in biological control of leaf spot on tall fescue (Festuca arundinacea), caused by Bipolaris sorokiniana, was investigated in vitro and in vivo. The filtrate of a broth culture of C3, with chitin as the carbon source, was separated into fractions. A high molecular-weight fraction (\u3e8 kDa) was chitinolytic and more inhibitory than a low-molecularweight, nonchitinolytic fraction to conidial germination and hyphal growth by B. sorokiniana and to leaf spot development. A protein fraction derived by ammonium sulfate precipitation and a chitinase fraction purified by chitin affinity chromatography also were chitinolytic and highly antifungal. The chitinolytic fractions caused swelling and vacuolation of conidia and discoloration, malformation, and degradation of germ tubes. When boiled, the chitinolytic fractions lost chitinase activity along with most of the antifungal properties. Two chitinase-deficient and two chitinase- reduced mutants of C3 were compared with the wild-type strain for inhibition of germination of B. sorokiniana conidia on tall fescue leaves and for suppression of leaf spot development in vivo. The mutants exhibited reduced antifungal activity and biocontrol efficacy, but did not lose all biocontrol activity. An aqueous extract of leaves colonized by wild-type C3 had higher chitinase activity than that of noncolonized leaves and was inhibitory to conidial germination. The addition of chitin to leaves along with the wild-type strain increased both chitinase and antifungal activity. The chitinase activity level of extracts from leaves colonized by a chitinase-deficient mutant of C3, with and without added chitin, was no higher than the background, and the extracts lacked antifungal activity. Chitinolysis appears to be one mechanism of biological control by strain C3, and it functions in concert with other mechanisms

Quantitative Characteristics of Gene Regulation by Small RNA

An increasing number of small RNAs (sRNAs) have been shown to regulate critical pathways in prokaryotes and eukaryotes. In bacteria, regulation by trans-encoded sRNAs is predominantly found in the coordination of intricate stress responses. The mechanisms by which sRNAs modulate expression of its targets are diverse. In common to most is the possibility that interference with the translation of mRNA targets may also alter the abundance of functional sRNAs. Aiming to understand the unique role played by sRNAs in gene regulation, we studied examples from two distinct classes of bacterial sRNAs in Escherichia coli using a quantitative approach combining experiment and theory. Our results demonstrate that sRNA provides a novel mode of gene regulation, with characteristics distinct from those of protein-mediated gene regulation. These include a threshold-linear response with a tunable threshold, a robust noise resistance characteristic, and a built-in capability for hierarchical cross-talk. Knowledge of these special features of sRNA-mediated regulation may be crucial toward understanding the subtle functions that sRNAs can play in coordinating various stress-relief pathways. Our results may also help guide the design of synthetic genetic circuits that have properties difficult to attain with protein regulators alone

Instability and topological robustness of Weyl semimetals against Coulomb interaction

There is a close connection between various new phenomena in Weyl semimetals and the existence of linear band crossings in the single particle description. We show, by a full self-consistent mean-field calculation, how this picture is modified in the presence of long-range Coulomb interactions. The chiral symmetry breaking occurs at strong enough interactions and the internode interband excitonic pairing channel is found to be significant, which determines the gap-opened band profile varying with interaction strength. Remarkably, in the resultant interacting phase, finite band Chern number jumps in the three-dimensional momentum space are retained, indicating the robustness of the topologically nontrivial features.Comment: 8 pages, 4 figures, accepted by Phys. Rev.

Coordination of gene expression with cell size enables Escherichia coli to efficiently maintain motility across conditions

To swim and navigate, motile bacteria synthesize a complex motility machinery involving flagella, motors, and a sensory system. A myriad of studies has elucidated the molecular processes involved, but less is known about the coordination of motility expression with cellular physiology: In Escherichia coli, motility genes are strongly up-regulated in nutrient-poor conditions compared to nutrient-replete conditions; yet a quantitative link to cellular motility has not been developed. Here, we systematically investigated gene expression, swimming behavior, cell growth, and available proteomics data across a broad spectrum of exponential growth conditions. Our results suggest that cells up-regulate the expression of motility genes at slow growth to compensate for reduction in cell size, such that the number of flagella per cell is maintained across conditions. The observed four or five flagella per cell is the minimum number needed to keep the majority of cells motile. This simple regulatory objective allows E. coli cells to remain motile across a broad range of growth conditions, while keeping the biosynthetic and energetic demands to establish and drive the motility machinery at the minimum needed. Given the strong reduction in flagella synthesis resulting from cell size increases at fast growth, our findings also provide a different physiological perspective on bacterial cell size control: A larger cell size at fast growth is an efficient strategy to increase the allocation of cellular resources to the synthesis of those proteins required for biomass synthesis and growth, while maintaining processes such as motility that are only needed on a per-cell basis

A Novel Mechanism of Transposon-Mediated Gene Activation

Transposable Insertion Sequences (IS elements) have been shown to provide various benefits to their hosts via gene activation or inactivation under stress conditions by appropriately inserting into specific chromosomal sites. Activation is usually due to derepression or introduction of a complete or partial promoter located within the element. Here we define a novel mechanism of gene activation by the transposon IS5 in Escherichia coli. The glycerol utilization operon, glpFK, that is silent in the absence of the cAMP-Crp complex, is activated by IS5 when inserted upstream of its promoter. High-level expression is nearly constitutive, only mildly dependent on glycerol, glucose, GlpR, and Crp, and allows growth at a rate similar to or more rapid than that of wild-type cells. Expression is from the glpFK promoter and dependent on (1) the DNA phase, (2) integration host factor (IHF), and (3) a short region at the 3′ end of IS5 harboring a permanent bend and an IHF binding site. The lacZYA operon is also subject to such activation in the absence of Crp. Thus, we have defined a novel mechanism of gene activation involving transposon insertion that may be generally applicable to many organisms

Conformational Transition Pathway in the Inhibitor Binding Process of Human Monoacylglycerol Lipase

Human monoacylglycerol lipase (MGL) catalyzes the hydrolysis of 2-arachidonoylglycerol to arachidonic and glycerol, which plays a pivotal role in the normal biological processes of brain. Co-crystal structure of the MGL in complex with its inhibitor, compound 1, shows that the helix α4 undergoes large-scale conformational changes in response to the compound 1 binding compared to the apo MGL. However, the detailed conformational transition pathway of the helix α4 in the inhibitor binding process of MGL has remained unclear. Here, conventional molecular dynamics (MD) and nudged elastic band (NEB) simulations were performed to explore the conformational transition pathway of the helix α4. Conventional MD simulations unveiled that the compound 1 induced the closed conformation of the active site of MGL, reduced the conformational flexibility of the helix α4, and elicited the large-scale conformational rearrangement of the helix α4, leading to the complete folding of the helix α4. Moreover, NEB simulations revealed that the conformational transition pathway of helix α4 underwent an almost 180° counter-clockwise rotation of the helix α4. Our computational results advance the structural and mechanistic understanding of the inhibitory mechanism

Characterization of the Stenotrophomonas maltophilia C3 antagonism system against Bipolaris sorokiniana on tall fescue

Stenotrophomonas maltophifia C3 inhibited the germination of conidia and the growth of germtubes of Bipolaris sorokiniana (Sacc.) Shoemaker on tall fescue (Festuca arundinacea Schreb.) leaves. It reduced the incidence and severity of leaf spot disease caused by B. sorokiniana under growth chamber and field conditions. Chitinolysis was found to be one mechanism of action. Transposon mutants of C3 that are chitinase deficient or have reduced chitinase production colonized grass to lower levels and provided reduced disease protection in comparison to the wild type. Chitinase activity on grass leaves and bean blossoms colonized by C3 was higher than on the non-colonized plant parts; the addition of chitin with C3 further increased chitinase activity. When a chitinase-minus mutant was applied to plant parts with or without chitin, no additional chitinase activity was detected over the controls. Extracts from C3-colonized plant parts had antifungal activity, and this property was related to chitinase activity. Chitinase production by C3 was induced in vitro by chitin or chitin-containing fungal cell walls. Chitinolytic fractions of C3 in broth cultures, that were partially purified by chitin affinity chromatography, were more antifungal than non-chitinolytic fractions. They also exhibited strong exochitinase and slight endochitinase activities. When the major chitinolytic fraction was subjected to SDS-PAGE, five proteins were revealed (25, 32, 48, 65 and 75 KDa). Only the N-terminal amino acid sequence of the 32-Kda protein showed homology to known bacterial chitinases. Five chitinase-active bands were detected when the gels were probed with a fluorescent chitin substrate. These bands appeared only as 32-Kda and 48-Kda proteins when re-electrophoresed in denaturing gels, suggesting that the smaller proteins were cleavage products of the larger proteins. Several methods were identified that increased disease control efficacy: application of C3 cells in combination with chitin, application of cells in combination with fluid from, a chitin-containing broth culture, and use of C3 cells induced to produced chitinase. Thus, improvements in biocontrol can be achieved with knowledge of the modes of action