The C8ORF38 homologue Sicily is a cytosolic chaperone for a mitochondrial complex I subunit

Mitochondrial complex I (CI) is an essential component in energy production through oxidative phosphorylation. Most CI subunits are encoded by nuclear genes, translated in the cytoplasm, and imported into mitochondria. Upon entry, they are embedded into the mitochondrial inner membrane. How these membrane-associated proteins cope with the hydrophilic cytoplasmic environment before import is unknown. In a forward genetic screen to identify genes that cause neurodegeneration, we identified sicily, the Drosophila melanogaster homologue of human C8ORF38, the loss of which causes Leigh syndrome. We show that in the cytoplasm, Sicily preprotein interacts with cytosolic Hsp90 to chaperone the CI subunit, ND42, before mitochondrial import. Loss of Sicily leads to loss of CI proteins and preproteins in both mitochondria and cytoplasm, respectively, and causes a CI deficiency and neurodegeneration. Our data indicate that cytosolic chaperones are required for the subcellular transport of ND42.     

Intrinsic microtubule GTP-cap dynamics in semi-confined systems: kinetochore–microtubule interface

In order to quantify the intrinsic dynamics associated with the tip of a GTP-cap under semi-confined conditions, such as those within a neuronal cone and at a kinetochore–microtubule interface, we propose a novel quantitative concept of critical nano local GTP-tubulin concentration (CNLC). A simulation of a rate constant of GTP-tubulin hydrolysis, under varying conditions based on this concept, generates results in the range of 0-420 s⁻¹. These results are in agreement with published experimental data, validating our model. The major outcome of this model is the prediction of 11 random and distinct outbursts of GTP hydrolysis per single layer of a GTP-cap. GTP hydrolysis is accompanied by an energy release and the formation of discrete expanding zones, built by less-stable, skewed GDP-tubulin subunits. We suggest that the front of these expanding zones within the walls of the microtubule represent soliton-like movements of local deformation triggered by energy released from an outburst of hydrolysis. We propose that these solitons might be helpful in addressing a long-standing question relating to the mechanism underlying how GTP-tubulin hydrolysis controls dynamic instability. This result strongly supports the prediction that large conformational movements in tubulin subunits, termed dynamic transitions, occur as a result of the conversion of chemical energy that is triggered by GTP hydrolysis (Satarić et al., Electromagn Biol Med 24:255–264, 2005). Although simple, the concept of CNLC enables the formulation of a rationale to explain the intrinsic nature of the “push-and-pull” mechanism associated with a kinetochore–microtubule complex. In addition, the capacity of the microtubule wall to produce and mediate localized spatio-temporal excitations, i.e., soliton-like bursts of energy coupled with an abundance of microtubules in dendritic spines supports the hypothesis that microtubule dynamics may underlie neural information processing including neurocomputation (Hameroff, J Biol Phys 36:71–93, 2010; Hameroff, Cognit Sci 31:1035–1045, 2007; Hameroff and Watt, J Theor Biol 98:549–561, 1982).     

Aspergillus Collagen-Like Genes (acl): Identification,Sequence Polymorphism,and Assessment for PCR-Based Pathogen Detection

The genus Aspergillus is a burden to public health due to its ubiquitous presence in the environment, its production of allergens, and wide demographic susceptibility among cystic fibrosis, asthmatic, and immunosuppressed patients. Current methods of detection of Aspergillus colonization and infection rely on lengthy morphological characterization or nonstandardized serological assays that are restricted to identifying a fungal etiology. Collagen-like genes have been shown to exhibit species-specific conservation across the noncollagenous regions as well as strain-specific polymorphism in the collagen-like regions. Here we assess the conserved region of the Aspergillus collagen-like (acl) genes and explore the application of PCR amplicon size-based discrimination among the five most common etiologic species of the Aspergillus genus, including Aspergillus fumigatus, A. flavus, A. nidulans, A. niger, and A. terreus. Genetic polymorphism and phylogenetic analysis of the aclF1 gene were additionally examined among the available strains. Furthermore, the applicability of the PCR-based assay to identification of these five species in cultures derived from sputum and bronchoalveolar fluid from 19 clinical samples was explored. Application of capillary electrophoresis on nanogels was additionally demonstrated to improve the discrimination between Aspergillus species. Overall, this study demonstrated that Aspergillus acl genes could be used as PCR targets to discriminate between clinically relevant Aspergillus species. Future studies aim to utilize the detection of Aspergillus acl genes in PCR and microfluidic applications to determine the sensitivity and specificity for the identification of Aspergillus colonization and invasive aspergillosis in immunocompromised subjects.     

Geomicrobiology of Pyrite (FeS2) Dissolution: Case Study at Iron Mountain,California

Geomicrobiology of pyrite weathering at Iron Mountain, CA, was investigated by molecular biological, surface chemical, surface structural, and solution chemical methods in both laboratory and field-based studies. Research focused at sites both within and peripheral to the ore-body. The acid-generating areas we have examined thus far at Iron Mountain (solution pH <1.0, temperature> 35 C) were populated by species other than Thiobacillus ferrooxidans . 16S rDNA bacterial sequence analysis and domain- and specieslevel oligonucleotide probe-based investigations confirmed the presence of planktonic Leptospirillum ferrooxidans and indicated the existence of other species apparently related to other newly described acidophilic chemolithotrophs. T. ferrooxidans was confined to relatively moderate environments (pH 2-3, 20-30 C) that were peripheral to the orebody. Dissolution rate measurements indicated that, per cell, attached and planktonic species contributed comparably in acid release. Surface colonization experiments in the laboratory and field indicated that attachment was specific to sulfides instead of to silicates, occurred in crystallographically preferred orientations, and, after cell division, resulted in a monolayer of cells at a maximum density of 8 X 106 cells cm2. In situ geochemical characterization throughout the year revealed that the microbial community that controlled acid generation varied and could be correlated with seasonal and spatial fluctuations in geochemical conditions.     

Regulation of insulin-regulated membrane aminopeptidase activity by its C-terminal domain

The development of inhibitors of insulin-regulated aminopeptidase (IRAP), a membrane-bound zinc metallopeptidase, is a promising approach for the discovery of drugs for the treatment of memory loss such as that associated with Alzheimer's disease. There is, however, no consensus in the literature about the mechanism by which inhibition occurs. Sequence alignments, secondary structure predictions, and homology models based on the structures of recently determined related metallopeptidases suggest that the extracellular region consists of four domains. Partial proteolysis and mass spectrometry reported here confirm some of the domain boundaries. We have produced purified recombinant fragments of human IRAP on the basis of these data and examined their kinetic and biochemical properties. Full-length extracellular constructs assemble as dimers with different nonoverlapping fragments dimerizing as well, suggesting an extended dimer interface. Only recombinant fragments containing domains 1 and 2 possess aminopeptidase activity and bind the radiolabeled hexapeptide inhibitor, angiotensin IV (Ang IV). However, fragments lacking domains 3 and 4 possess reduced activity, although they still bind a range of inhibitors with the same affinity as longer fragments. In the presence of Ang IV, IRAP is resistant to proteolysis, suggesting significant conformational changes occur upon binding of the inhibitor. We show that IRAP has a second Zn(2+) binding site, not associated with the catalytic region, which is lost upon binding Ang IV. Modulation of activity caused by domains 3 and 4 is consistent with a conformational change regulating access to the active site of IRAP.     

Evidence for 20th century climate warming and wetland drying in the North American Prairie Pothole Region

The Prairie Pothole Region (PPR) of North America is a globally important resource that provides abundant and valuable ecosystem goods and services in the form of biodiversity, groundwater recharge, water purification, flood attenuation, and water and forage for agriculture. Numerous studies have found these wetlands, which number in the millions, to be highly sensitive to climate variability . Here, we compare wetland conditions between two 30‐year periods (1946–1975; 1976–2005) using a hindcast simulation approach to determine if recent climate warming in the region has already resulted in changes in wetland condition. Simulations using the WETLANDSCAPE model show that 20th century climate change may have been sufficient to have a significant impact on wetland cover cycling. Modeled wetlands in the PPR's western Canadian prairies show the most dramatic effects: a recent trend toward shorter hydroperiods and less dynamic vegetation cycles, which already may have reduced the productivity of hundreds of wetland ‐ dependent species.     

Effects of Sexual Dimorphism and Landscape Composition on the Trophic Behavior of Greater Prairie-Chicken

Partitioning of ecological niche is expected in lekking species that show marked sexual size dimorphism as a consequence of sex-specific ecological constraints. However, niche partitioning is uncertain in species with moderate sexual dimorphism. In addition, the ecological niche of a species may also be affected by landscape composition; particularly, agricultural fragmentation may greatly influence the trophic behavior of herbivores. We studied trophic niche variation in Greater Prairie-Chickens (Tympanuchus cupido), a grouse species that shows moderate sex-dimorphism. Greater Prairie-Chickens are native to tallgrass prairies of North America, although populations persist in less natural mosaics of cropland and native habitats. We used stable isotope analysis of carbon and nitrogen in blood, claws and feathers to assess seasonal differences in trophic niche breadth and individual specialization between male and female Greater Prairie-Chickens, and between birds living in continuous and fragmented landscapes. We found that females showed broader niches and higher individual specialization than males, especially in winter and autumn. However, differences between females and males were smaller in spring when birds converge at leks, suggesting that females and males may exhibit similar feeding behaviors during the lekking period. In addition, we found that birds living in native prairies showed greater annual trophic variability than conspecifics in agricultural mosaic landscapes. Native habitats may provide greater dietary diversity, resulting in greater diversity of feeding strategies.