Pseudomonas aeruginosa Homoserine Lactone Activates Store-operated cAMP and Cystic Fibrosis Transmembrane Regulator-dependent Cl? Secretion by Human Airway Epithelia

The ubiquitous bacterium Pseudomonas aeruginosa frequently causes hospital-acquired infections. P. aeruginosa also infects the lungs of cystic fibrosis (CF) patients and secretes N-(3-oxo-dodecanoyl)-S-homoserine lactone (3O-C12) to regulate bacterial gene expression critical for P. aeruginosa persistence. In addition to its effects as a quorum-sensing gene regulator in P. aeruginosa, 3O-C12 elicits cross-kingdom effects on host cell signaling leading to both pro- or anti-inflammatory effects. We find that in addition to these slow effects mediated through changes in gene expression, 3O-C12 also rapidly increases Cl⁻ and fluid secretion in the cystic fibrosis transmembrane regulator (CFTR)-expressing airway epithelia. 3O-C12 does not stimulate Cl⁻ secretion in CF cells, suggesting that lactone activates the CFTR. 3O-C12 also appears to directly activate the inositol trisphosphate receptor and release Ca²⁺ from the endoplasmic reticulum (ER), lowering [Ca²⁺] in the ER and thereby activating the Ca²⁺-sensitive ER signaling protein STIM1. 3O-C12 increases cytosolic [Ca²⁺] and, strikingly, also cytosolic [cAMP], the known activator of CFTR. Activation of Cl⁻ current by 3O-C12 was inhibited by a cAMP antagonist and increased by a phosphodiesterase inhibitor. Finally, a Ca²⁺ buffer that lowers [Ca²⁺] in the ER similar to the effect of 3O-C12 also increased cAMP and I_Cl. The results suggest that 3O-C12 stimulates CFTR-dependent Cl⁻ and fluid secretion in airway epithelial cells by activating the inositol trisphosphate receptor, thus lowering [Ca²⁺] in the ER and activating STIM1 and store-operated cAMP production. In CF airways, where CFTR is absent, the adaptive ability to rapidly flush the bacteria away is compromised because the lactone cannot affect Cl⁻ and fluid secretion.     

Variation in floral sex allocation in Polygonatum odoratum (Liliaceae)

BACKGROUND AND AIMS: It is well known that resource allocation to male and female functions can be highly variable in hermaphroditic plants. The purpose of this study was to investigate variations in sexual investment at different levels (flower, plant and population) in Polygonatum odoratum, a plant with sequentially opening flowers. METHODS: Pollen and ovule production in base, middle and top flowers of P. odoratum flowering shoots from two natural populations were quantified. Plant measurements of phenotypic and functional gender were calculated in both populations. Total leaf number was used to investigate the relationship between gender assessments and plant size. KEY RESULTS: Pollen and ovule production varied depending on flower position, although the precise pattern differed between both studied populations; only investment in female floral function decreased markedly from base to top flowers in both populations. The frequency distribution of phenotypic gender and their relationship with plant size differed between populations. Phenotypic and functional gender were correlated in both populations. CONCLUSIONS: Sexual investment in P. odoratum has shown a marked variability within plants, among plants, and between populations, which confirms the importance of analysing sex expression in plants of this type. Differences in relative investment in male and female components (phenotypic gender) are reflected in the functional gender and it would be expected that the evolution of sexual specialization in Polygonatum odoratum would be promoted.     

A change from phalanx to guerrilla growth form is an effective strategy to acclimate to sedimentation in a wetland sedge species Carex brevicuspis (Cyperaceae)

The rhizomatous sedge Carex brevicuspis can produce clumping ramets from shortened rhizomes (phalanx) and spreading ramets from elongated rhizomes (guerrilla) to form a combined clonal growth form. In this paper, changes in clonal growth and biomass allocation pattern of C. brevicuspis in response to sedimentation were studied. Four sedimentation depths (0, 3, 6, and 9 cm) were applied to 48 ramets in a randomized block design. Plants were harvested after 20 weeks. With increasing sedimentation depth, the proportion of spreading ramets to total ramets increased from 19.6% in 0 cm to 92.9% in 9 cm sedimentation treatments, whereas that of clumping ramets decreased from 80.4% to 7.1%, indicating a change of clonal growth form from phalanx to guerrilla as a response to sedimentation. With increasing sedimentation depth, biomass allocation to shoots and roots did not change, but rhizome mass ratio increased from 2.7% in 0 cm to 7.2% in 9 cm sedimentation treatments, suggesting that production of long rhizomes changes biomass allocation pattern. The results show that plasticity of clonal growth forms, by which more spreading ramets are produced, is an effective strategy to avoid sedimentation stress under our experimental conditions.     

三种温带树种非结构性碳水化合物的分配

树体中的非结构性碳水化合物(NSC)浓度、含量及其分配反映了树木整体的碳供应状况, 是决定树木生长和存活的关键因子, 也是构建树木碳平衡模型的关键参数。温带树种的NSC尚缺乏系统研究。该文测定了特性各异的3种温带树种在生长盛期的NSC及其组分的浓度和含量以及分配格局的种间种内变异。结果表明, NSC及其组分的浓度在树种和组织之间差异显著, 可溶性糖、淀粉和总NSC浓度分别在0.65-8.45、1.96-5.95和3.00-13.90 g·100 g^-1 DM之间波动。NSC及其组分含量的大小依次为: 兴安落叶松(Larix gmelinii) >蒙古栎( Quercus mongolica) >红松( Pinus koraiensis), 其中叶和根中的浓度较高。树干中的NSC及其组分浓度的纵向变化不显著, 但其心材与边材之间的浓度差异却随树种和NSC组分而异, 表现为心边材的可溶性糖浓度差异不显著, 但其淀粉和总NSC浓度差异显著。不同直径根系的NSC及其组分浓度在2种针叶树种中差异不显著, 但在蒙古栎中差异显著。蒙古栎将可溶性糖主要投资到地上生长, 而2种针叶树将更多的可溶性糖投资到根系生长。淀粉的主要储存库为树干, 其在树体内的分布格局与可溶性糖正相反, 因而使总NSC在树根和树枝中的分配趋于较平衡状态。在树干中, 除了2种针叶树的可溶性糖库以边材为主外, 心材是淀粉和总NSC的主要储存库。在树根中, 粗根是NSC及其组分的优势储存库。该研究中3种温带树种的NSC及其组分的浓度和含量的种间和种内变化, 反映了这些树种的生长对策和体内碳源汇强度的差异。     

Protein-protein interactions in assembly of lipoic acid on the 2-oxoacid dehydrogenases of aerobic metabolism

Lipoic acid is a covalently attached cofactor essential for the activity of 2-oxoacid dehydrogenases and the glycine cleavage system. In the absence of lipoic acid modification, the dehydrogenases are inactive, and aerobic metabolism is blocked. In Escherichia coli, two pathways for the attachment of lipoic acid exist, a de novo biosynthetic pathway dependent on the activities of the LipB and LipA proteins and a lipoic acid scavenging pathway catalyzed by the LplA protein. LipB is responsible for octanoylation of the E2 components of 2-oxoacid dehydrogenases to provide the substrates of LipA, an S-adenosyl-L-methionine radical enzyme that inserts two sulfur atoms into the octanoyl moiety to give the active lipoylated dehydrogenase complexes. We report that the intact pyruvate and 2-oxoglutarate dehydrogenase complexes specifically copurify with both LipB and LipA. Proteomic, genetic, and dehydrogenase activity data indicate that all of the 2-oxoacid dehydrogenase components are present. In contrast, LplA, the lipoate protein ligase enzyme of lipoate salvage, shows no interaction with the 2-oxoacid dehydrogenases. The interaction is specific to the dehydrogenases in that the third lipoic acid-requiring enzyme of Escherichia coli, the glycine cleavage system H protein, does not copurify with either LipA or LipB. Studies of LipB interaction with engineered variants of the E2 subunit of 2-oxoglutarate dehydrogenase indicate that binding sites for LipB reside both in the lipoyl domain and catalytic core sequences. We also report that LipB forms a very tight, albeit noncovalent, complex with acyl carrier protein. These results indicate that lipoic acid is not only assembled on the dehydrogenase lipoyl domains but that the enzymes that catalyze the assembly are also present "on site."     

Disturbance and climate effects on carbon stocks and fluxes across Western Oregon USA

We used a spatially nested hierarchy of field and remote‐sensing observations and a process model, Biome‐BGC, to produce a carbon budget for the forested region of Oregon, and to determine the relative influence of differences in climate and disturbance among the ecoregions on carbon stocks and fluxes. The simulations suggest that annual net uptake (net ecosystem production (NEP)) for the whole forested region (8.2 million hectares) was 13.8 Tg C (168 g C m^?2 yr^?1), with the highest mean uptake in the Coast Range ecoregion (226 g C m^?2 yr^?1), and the lowest mean NEP in the East Cascades (EC) ecoregion (88 g C m^?2 yr^?1). Carbon stocks totaled 2765 Tg C (33 700 g C m^?2), with wide variability among ecoregions in the mean stock and in the partitioning above‐ and belowground. The flux of carbon from the land to the atmosphere that is driven by wildfire was relatively low during the late 1990s (～0.1 Tg C yr^?1), however, wildfires in 2002 generated a much larger C source (～4.1 Tg C). Annual harvest removals from the study area over the period 1995–2000 were ～5.5 Tg C yr^?1. The removals were disproportionately from the Coast Range, which is heavily managed for timber production (approximately 50% of all of Oregon's forest land has been managed for timber in the past 5 years). The estimate for the annual increase in C stored in long‐lived forest products and land fills was 1.4 Tg C yr^?1. Net biome production (NBP) on the land, the net effect of NEP, harvest removals, and wildfire emissions indicates that the study area was a sink (8.2 Tg C yr^?1). NBP of the study area, which is the more heavily forested half of the state, compensated for ～52% of Oregon's fossil carbon dioxide emissions of 15.6 Tg C yr^?1 in 2000. The Biscuit Fire in 2002 reduced NBP dramatically, exacerbating net emissions that year. The regional total reflects the strong east–west gradient in potential productivity associated with the climatic gradient, and a disturbance regime that has been dominated in recent decades by commercial forestry.     

Construction of hybrid photosynthetic units using peripheral and core antennae from two different species of photosynthetic bacteria: detection of the energy transfer from bacteriochlorophyll a in LH2 to bacteriochlorophyll b in LH1

Typical purple bacterial photosynthetic units consist of supra-molecular arrays of peripheral (LH2) and core (LH1-RC) antenna complexes. Recent atomic force microscopy pictures of photosynthetic units in intact membranes have revealed that the architecture of these units is variable (Scheuring et al. (2005) Biochim Bhiophys Acta 1712:109–127). In this study, we describe methods for the construction of heterologous photosynthetic units in lipid-bilayers from mixtures of purified LH2 (from Rhodopseudomonas acidophila) and LH1-RC (from Rhodopseudomonas viridis) core complexes. The architecture of these reconstituted photosynthetic units can be varied by controlling ratio of added LH2 to core complexes. The arrangement of the complexes was visualized by electron-microscopy in combination with Fourier analysis. The regular trigonal array of the core complexes seen in the native photosynthetic membrane could be regenerated in the reconstituted membranes by temperature cycling. In the presence of added LH2 complexes, this trigonal symmetry was replaced with orthorhombic symmetry. The small lattice lengths for the latter suggest that the constituent unit of the orthorhombic lattice is the LH2. Fluorescence and fluorescence-excitation spectroscopy was applied to the set of the reconstituted membranes prepared with various proportions of LH2 to core complexes. Remarkably, even though the LH2 complexes contain bacteriochlorophyll a, and the core complexes contain bacteriochlorophyll b, it was possible to demonstrate energy transfer from LH2 to the core complexes. These experiments provide a first step along the path toward investigating how changing the architecture of purple bacterial photosynthetic units affects the overall efficiency of light-harvesting.