Effect of Levels of Acetate on the Mevalonate Pathway of Borrelia burgdorferi

Borrelia burgdorferi, the agent of Lyme disease, is a spirochetal pathogen with limited metabolic capabilities that survives under highly disparate host-specific conditions. However, the borrelial genome encodes several proteins of the mevalonate pathway (MP) that utilizes acetyl-CoA as a substrate leading to intermediate metabolites critical for biogenesis of peptidoglycan and post-translational modifications of proteins. In this study, we analyzed the MP and contributions of acetate in modulation of adaptive responses in B. burgdorferi. Reverse-transcription PCR revealed that components of the MP are transcribed as individual open reading frames. Immunoblot analysis using monospecific sera confirmed synthesis of members of the MP in B. burgdorferi. The rate-limiting step of the MP is mediated by HMG-CoA reductase (HMGR) via conversion of HMG-CoA to mevalonate. Recombinant borrelial HMGR exhibited a Km value of 132 µM with a Vmax of 1.94 µmol NADPH oxidized minute−1 (mg protein)−1 and was inhibited by statins. Total protein lysates from two different infectious, clonal isolates of B. burgdorferi grown under conditions that mimicked fed-ticks (pH 6.8/37°C) exhibited increased levels of HMGR while other members of the MP were elevated under unfed-tick (pH 7.6/23°C) conditions. Increased extra-cellular acetate gave rise to elevated levels of MP proteins along with RpoS, CsrABb and their respective regulons responsible for mediating vertebrate host-specific adaptation. Both lactone and acid forms of two different statins inhibited growth of B. burgdorferi strain B31, while overexpression of HMGR was able to partially overcome that inhibition. In summary, these studies on MP and contributions of acetate to host-specific adaptation have helped identify potential metabolic targets that can be manipulated to reduce the incidence of Lyme disease

BosR (BB0647) Controls the RpoN-RpoS Regulatory Pathway and Virulence Expression in Borrelia burgdorferi by a Novel DNA-Binding Mechanism

In Borrelia burgdorferi (Bb), the Lyme disease spirochete, the alternative σ factor σ54 (RpoN) directly activates transcription of another alternative σ factor, σS (RpoS) which, in turn, controls the expression of virulence-associated membrane lipoproteins. As is customary in σ54-dependent gene control, a putative NtrC-like enhancer-binding protein, Rrp2, is required to activate the RpoN-RpoS pathway. However, recently it was found that rpoS transcription in Bb also requires another regulator, BosR, which was previously designated as a Fur or PerR homolog. Given this unexpected requirement for a second activator to promote σ54-dependent gene transcription, and the fact that regulatory mechanisms among similar species of pathogenic bacteria can be strain-specific, we sought to confirm the regulatory role of BosR in a second virulent strain (strain 297) of Bb. Indeed, BosR displayed the same influence over lipoprotein expression and mammalian infectivity for strain Bb 297 that were previously noted for Bb strain B31. We subsequently found that recombinant BosR (rBosR) bound to the rpoS gene at three distinct sites, and that binding occurred despite the absence of consensus Fur or Per boxes. This led to the identification of a novel direct repeat sequence (TAAATTAAAT) critical for rBosR binding in vitro. Mutations in the repeat sequence markedly inhibited or abolished rBosR binding. Taken together, our studies provide new mechanistic insights into how BosR likely acts directly on rpoS as a positive transcriptional activator. Additional novelty is engendered by the facts that, although BosR is a Fur or PerR homolog and it contains zinc (like Fur and PerR), it has other unique features that clearly set it apart from these other regulators. Our findings also have broader implications regarding a previously unappreciated layer of control that can be involved in σ54–dependent gene regulation in bacteria

A unified model for BAM function that takes into account type Vc secretion and species differences in BAM composition

Transmembrane proteins in the outer membrane of Gram-negative bacteria are almost exclusively β-barrels. They are inserted into the outer membrane by a conserved and essential protein complex called the BAM (for β-barrel assembly machinery). In this commentary, we summarize current research into the mechanism of this protein complex and how it relates to type V secretion. Type V secretion systems are autotransporters that all contain a β-barrel transmembrane domain inserted by BAM. In type Vc systems, this domain is a homotrimer. We argue that none of the current models are sufficient to explain BAM function particularly regarding type Vc secretion. We also find that current models based on the well-studied model system Escherichia coli mostly ignore the pronounced differences in BAM composition between different bacterial species. We propose a more holistic view on how all OMPs, including autotransporters, are incorporated into the lipid bilayer

Land Conservation Can Mitigate Freshwater Ecosystem Services Degradation Due to Climate Change in a Semiarid Catchment: The Case of the Portneuf River Catchment, Idaho, USA

There is increasing evidence of environmental change impacts on freshwater ecosystem services especially through land use and climate change. However, little is known about how land conservation could help mitigate adverse water-sustainability impacts. In this paper, we utilized the InVEST tool and the Residual Trends method to assess the joint effects and relative contributions of climate change and land conservation on freshwater ecosystem services in the Portneuf River catchment in Idaho, USA. We developed five hypothesized scenarios regarding gain and loss in the enrollment of Conservation Reserve Program (CRP), the largest agricultural land-retirement program in the U.S., plus riparian buffer and assessed their interactions with climate change. Results suggest that the realized water yield in the Portneuf River catchment would possibly be 56% less due to climate change and 24% less due to the decline of CRP enrollment. On the contrary, if CRP enrollment is promoted by ~30% and riparian buffer protection is implemented, the water supply reduction in the year 2050 could be changed from 56% to 26%, the total phosphorus (TP) and total nitrogen (TN) export would be reduced by 10% and 11%, and the total suspended sediment (TSS) reduced by 17%. This study suggests that increasing implementation of the CRP would likely preserve key freshwater ecosystem services and assist proactive mitigation, especially for semiarid regions vulnerable to changing climate conditions

Nível de dano econômico como critério para controle de picão-preto em soja Threshold level as a criterion for beggartick control in soybeans

A adoção do conceito de nível de dano econômico (NDE) no manejo de plantas daninhas avalia suas populações de modo que medidas de controle sejam implementadas somente quando as infestações superarem os NDEs. O objetivo deste trabalho foi definir o NDE para infestações mistas das espécies Bidens pilosa e Bidens subalternans na cultura da soja. Foram conduzidos experimentos em campo em dois locais: Passo Fundo e Eldorado do Sul, RS. Os tratamentos constaram de densidades mistas das espécies de picão-preto e semeadura da soja aos 3, 7 e 11 dias após dessecação da cobertura vegetal. Obtiveram-se valores para NDE variáveis de 0,4 a 33 plantas m-2. Foram verificadas respostas diferentes nas relações de interferência cultura-plantas daninhas entre os locais estudados. O atraso na semeadura da soja em relação à dessecação da cobertura vegetal incrementa o grau de interferência de picão-preto na cultura. Detectou-se que aumentos na perda de rendimento por unidade de planta daninha, no potencial de rendimento da cultura, no valor do produto colhido e na eficiência do herbicida diminuem os valores de NDE, tornando potencialmente mais econômico o controle; já aumento no custo do controle das plantas daninhas faz elevar os NDEs. Constatou-se que as sementes produzidas por plantas não-controladas, ocorrendo em densidades abaixo do NDE, comprometem a adoção da tomada de decisão de controle com base neste critério.<br>Adoption of the threshold level (TL) concept for weed management evaluates weed populations to implement control measures only when infestations overcome the TL. The objective of this research was to define TL for mixed infestations of Bidens pilosa and Bidens subalternans in the soybean crop. Field experiments were conducted in Passo Fundo and Eldorado do Sul, RS, Brazil. Treatments consisted of mixed densities of the beggartick species and soybean seeding at 3, 7, and 11 days after cover crop desiccation. TL variable values were obtained from 0.4 to 33 beggartick plants m-2. Different responses were obtained for crop-weed interference relations between the two locations evaluated. Delaying soybean seeding in relation to cover crop desiccation increases beggartick interference in this crop. Increases in yield loss per weed unit, in crop yield potential, harvested product value and herbicide efficiency were found to decrease TL values, making control potentially more economic. On the other hand, higher weed control cost increases TL values. It was also verified that seeds produced by non-controlled weeds below TL could compromise adoption of control decision based on this criterion