Pathogen protein modularity enables elaborate mimicry of a host phosphatase.

Pathogens produce diverse effector proteins to manipulate host cellular processes. However, how functional diversity is generated in an effector repertoire is poorly understood. Many effectors in the devastating plant pathogen Phytophthora contain tandem repeats of the "(L)WY" motif, which are structurally conserved but variable in sequences. Here, we discovered a functional module formed by a specific (L)WY-LWY combination in multiple Phytophthora effectors, which efficiently recruits the serine/threonine protein phosphatase 2A (PP2A) core enzyme in plant hosts. Crystal structure of an effector-PP2A complex shows that the (L)WY-LWY module enables hijacking of the host PP2A core enzyme to form functional holoenzymes. While sharing the PP2A-interacting module at the amino terminus, these effectors possess divergent C-terminal LWY units and regulate distinct sets of phosphoproteins in the host. Our results highlight the appropriation of an essential host phosphatase through molecular mimicry by pathogens and diversification promoted by protein modularity in an effector repertoire.

Chemical reaction induced carrier localization in nanometer-thin Al/Ru, Al/Co, and Al/Mo superlattices.

It is well-known that the electrical conductivity of a metallic film reduces dramatically when the film becomes very thin. This effect is mainly attributed to surface scattering of the conducting carriers. In a multilayer structure, interface scattering also reduces the conductance, but chemical reactions at the interfaces can have equal or bigger effects. The extent of chemically induced carrier localization at the metallic interfaces has not been explored or reported. We have grown superlattices consisting of nm-thin, alternating Al and transition-metal layers (Al/Ru, Al/Co and Al/Mo) by magnetron sputtering, and measured the electrical conductance of the superlattices in-situ during the growth. We observed a sharp conductance drop at the start of each transition metal layer and a pause in conduction increase at the start of each Al layer, neither of which is predicted by the surface scattering model. We show that these abnormal conductance changes can be explained by localization of Al free carriers at the interfaces to facilitate the formation of intermetallic bonds. The magnitude of the measured conductance drops suggests that one monolayer of compound is formed at each interface at room temperature. Annealing the superlattices to 300 °C caused a modest decrease in conductance, attributed to further chemical reactions. In contrast, a superlattice involving two fully miscible transition metals, Ru and Co, exhibited no carrier localization, resulting in a conductance more than three times that of superlattices containing Al layers.

The RXLR Effector PcAvh1 Is Required for Full Virulence of Phytophthora capsici.

Plant pathogens employ diverse secreted effector proteins to manipulate host physiology and defense in order to foster diseases. The destructive Phytophthora pathogens encode hundreds of cytoplasmic effectors, which are believed to function inside the plant cells. Many of these cytoplasmic effectors contain the conserved N-terminal RXLR motif. Understanding the virulence function of RXLR effectors will provide important knowledge of Phytophthora pathogenesis. Here, we report the characterization of RXLR effector PcAvh1 from the broad-host range pathogen Phytophthora capsici. Only expressed during infection, PcAvh1 is quickly induced at the early infection stages. CRISPR/Cas9-knockout of PcAvh1 in P. capsici severely impairs virulence while overexpression enhances disease development in Nicotiana benthamiana and bell pepper, demonstrating that PcAvh1 is an essential virulence factor. Ectopic expression of PcAvh1 induces cell death in N. benthamiana, tomato, and bell pepper. Using yeast two-hybrid screening, we found that PcAvh1 interacts with the scaffolding subunit of the protein phosphatase 2A (PP2Aa) in plant cells. Virus-induced gene silencing of PP2Aa in N. benthamiana attenuates resistance to P. capsici and results in dwarfism, suggesting that PP2Aa regulates plant immunity and growth. Collectively, these results suggest that PcAvh1 contributes to P. capsici infection, probably through its interaction with host PP2Aa.

Small RNAs regulate plant responses to filamentous pathogens.

Small RNAs are central players of RNA silencing in eukaryotes. These short RNA molecules (20-25 nucleotides in length) repress target gene expression based on sequence complementarity. While small RNAs are well-known for their essential function in regulating growth and development, recent research has revealed that they also influence plant immunity. Extensive changes in small RNA accumulation have been observed during infection. This review focuses on specific small RNA changes that are involved in plant responses to filamentous eukaryotic pathogens including fungi and oomycetes. We describe how changes in small RNA accumulation influence plant immunity and summarize the cellular processes affected by these small RNAs. In particular, we discuss secondary small interfering RNAs that directly modulate the expression of defense-related genes.

A Polysaccharide Derived from a Trichosporon sp. Culture Strongly Primes Plant Resistance to Viruses.

Plant viruses cause devastating diseases in plants, yet no effective viricide is available for agricultural application. We screened cultured filtrates derived from various soil microorganisms cultured in vegetable broth that enhanced plant viral resistance. A cultured filtrate, designated F8 culture filtrate, derived from a fungus belonging to the genus Trichosporon, induced strong resistance to various viruses on different plants. Our inoculation assay found the infection rate of Tobacco mosaic virus (TMV)-inoculated Nicotiana benthamiana with F8 culture filtrate pretreatment may decrease to 0%, whereas salicylic acid (SA)-pretreated N. benthamiana attenuated TMV-caused symptoms but remained 100% infected. Tracking Tobacco mosaic virus tagged with green fluorescence protein in plants revealed pretreatment with F8 culture filtrate affected the initial establishment of the virus infection. From F8 culture filtrate, we identified a previously unknown polysaccharide composed of D-mannose, D-galactose, and D-glucose in the ratio 1.0:1.2:10.0 with a α-D-1,4-glucan linkage to be responsible for the induction of plant resistance against viruses through priming of SA-governed immune-responsive genes. Notably, F8 culture filtrate only triggered local defense but was much more effective than conventional SA-mediated systematic acquired resistance. Our finding revealed that microbial cultured metabolites provided a rich source for identification of potent elicitors in plant defense.

Effects of nanoscale surface roughness on the resistivity of ultrathin epitaxial copper films.

The knowledge on the influence of surface roughness and the electron-phonon (el-ph) interaction on electrical transport properties of nanoscale metal films is important from both fundamental and technological points of view. Here we report a study of the temperature dependent electron transport properties of nanoscale copper films by measuring temperature dependent electrical resistivity with thickness ranging from 4 to 500 nm. We show that the residual resistivity, which is temperature independent, can be described quantitatively using both measured vertical surface root-mean-square roughness and lateral correlation length in the nanoscale, with no adjustable parameter, by a recent quasi-classical model developed by Chatterjee and Meyerovich (2010 Phys. Rev. B 81 245409-10). We also demonstrate that the temperature dependent component of the resistivity can be described using the Bloch-Grüneisen equation with a thickness dependent el-ph coupling constant and a thickness dependent Debye temperature. We show that the increase of the el-ph coupling constant with the decrease of film thickness gives rise to an enhancement of the temperature dependent component of the resistivity.

Ultrastructural differences impact cilia shape and external exposure across cell classes in the visual cortex.

A primary cilium is a membrane-bound extension from the cell surface that contains receptors for perceiving and transmitting signals that modulate cell state and activity. Primary cilia in the brain are less accessible than cilia on cultured cells or epithelial tissues because in the brain they protrude into a deep, dense network of glial and neuronal processes. Here, we investigated cilia frequency, internal structure, shape, and position in large, high-resolution transmission electron microscopy volumes of mouse primary visual cortex. Cilia extended from the cell bodies of nearly all excitatory and inhibitory neurons, astrocytes, and oligodendrocyte precursor cells (OPCs) but were absent from oligodendrocytes and microglia. Ultrastructural comparisons revealed that the base of the cilium and the microtubule organization differed between neurons and glia. Investigating cilia-proximal features revealed that many cilia were directly adjacent to synapses, suggesting that cilia are poised to encounter locally released signaling molecules. Our analysis indicated that synapse proximity is likely due to random encounters in the neuropil, with no evidence that cilia modulate synapse activity as would be expected in tetrapartite synapses. The observed cell class differences in proximity to synapses were largely due to differences in external cilia length. Many key structural features that differed between neuronal and glial cilia influenced both cilium placement and shape and, thus, exposure to processes and synapses outside the cilium. Together, the ultrastructure both within and around neuronal and glial cilia suggest differences in cilia formation and function across cell types in the brain.

Nanometer-scale views of visual cortex reveal anatomical features of primary cilia poised to detect synaptic spillover.

A primary cilium is a thin membrane-bound extension off a cell surface that contains receptors for perceiving and transmitting signals that modulate cell state and activity. While many cell types have a primary cilium, little is known about primary cilia in the brain, where they are less accessible than cilia on cultured cells or epithelial tissues and protrude from cell bodies into a deep, dense network of glial and neuronal processes. Here, we investigated cilia frequency, internal structure, shape, and position in large, high-resolution transmission electron microscopy volumes of mouse primary visual cortex. Cilia extended from the cell bodies of nearly all excitatory and inhibitory neurons, astrocytes, and oligodendrocyte precursor cells (OPCs), but were absent from oligodendrocytes and microglia. Structural comparisons revealed that the membrane structure at the base of the cilium and the microtubule organization differed between neurons and glia. OPC cilia were distinct in that they were the shortest and contained pervasive internal vesicles only occasionally observed in neuron and astrocyte cilia. Investigating cilia-proximal features revealed that many cilia were directly adjacent to synapses, suggesting cilia are well poised to encounter locally released signaling molecules. Cilia proximity to synapses was random, not enriched, in the synapse-rich neuropil. The internal anatomy, including microtubule changes and centriole location, defined key structural features including cilium placement and shape. Together, the anatomical insights both within and around neuron and glia cilia provide new insights into cilia formation and function across cell types in the brain.