Metabolic Labeling of Caenorhabditis elegans Primary Embryonic Cells with Azido-Sugars as a Tool for Glycoprotein Discovery

Glycobiology research with Caenorhabditis elegans (C. elegans) has benefitted from the numerous genetic and cell biology tools available in this system. However, the lack of a cell line and the relative inaccessibility of C. elegans somatic cells in vivo have limited the biochemical approaches available in this model. Here we report that C. elegans primary embryonic cells in culture incorporate azido-sugar analogs of N-acetylgalactosamine (GalNAc) and N-acetylglucosamine (GlcNAc), and that the labeled glycoproteins can be analyzed by mass spectrometry. By using this metabolic labeling approach, we have identified a set of novel C. elegans glycoprotein candidates, which include several mitochondrially-annotated proteins. This observation was unexpected given that mitochondrial glycoproteins have only rarely been reported, and it suggests that glycosylation of mitochondrially-annotated proteins might occur more frequently than previously thought. Using independent experimental strategies, we validated a subset of our glycoprotein candidates. These include a mitochondrial, atypical glycoprotein (ATP synthase α-subunit), a predicted glycoprotein (aspartyl protease, ASP-4), and a protein family with established glycosylation in other species (actin). Additionally, we observed a glycosylated isoform of ATP synthase α-subunit in bovine heart tissue and a primate cell line (COS-7). Overall, our finding that C. elegans primary embryonic cells are amenable to metabolic labeling demonstrates that biochemical studies in C. elegans are feasible, which opens the door to labeling C. elegans cells with other radioactive or azido-substrates and should enable the identification of additional post-translationally modified targets and analysis of the genes required for their modification using C. elegans mutant libraries.     

Genome sequence of the homoacetogenic bacterium Holophaga foetida type strain (TMBS4(T))

Holophaga foetida Liesack et al. 1995 is a member of the phylum Acidobacteria and is of interest for its ability to anaerobically degrade aromatic compounds and for its production of volatile sulfur compounds through a unique pathway. The genome of H. foetida strain TMBS4(T) is the first to be sequenced for a representative of the class Holophagae. Here we describe the features of this organism, together with the complete genome sequence (improved high quality draft), and annotation. The 4,127,237 bp long chromosome with its 3,615 protein-coding and 57 RNA genes is a part of the Genomic Encyclopedia of Bacteria and Archaea project.     

Complete genome sequence of Dehalogenimonas lykanthroporepellens type strain (BL-DC-9(T)) and comparison to "Dehalococcoides" strains

Dehalogenimonas lykanthroporepellens is the type species of the genus Dehalogenimonas, which belongs to a deeply branching lineage within the phylum Chloroflexi. This strictly anaerobic, mesophilic, non spore-forming, Gram-negative staining bacterium was first isolated from chlorinated solvent contaminated groundwater at a Superfund site located near Baton Rouge, Louisiana, USA. D. lykanthroporepellens was of interest for genome sequencing for two reasons: (a) an unusual ability to couple growth with reductive dechlorination of environmentally important polychlorinated aliphatic alkanes and (b) a phylogenetic position that is distant from previously sequenced bacteria. The 1,686,510 bp circular chromosome of strain BL-DC-9(T) contains 1,720 predicted protein coding genes, 47 tRNA genes, a single large subunit rRNA (23S-5S) locus, and a single, orphan, small subunit rRNA (16S) locus.     

Primate origins,human origins,and the end of higher taxa

When people learn that I study human evolution and we start talking about it, they sometimes ask me, “How long ago did the first humans live?” My answer is usually another question: “What do you mean by 'humans'?” That response seems as baffling and wrong‐headed to them as their question seems to me, and it usually takes us a while to straighten things out. © 2012 Wiley Periodicals, Inc.     

Reference genome sequence of the model plant Setaria

We generated a high-quality reference genome sequence for foxtail millet (Setaria italica). The ～400-Mb assembly covers ～80% of the genome and >95% of the gene space. The assembly was anchored to a 992-locus genetic map and was annotated by comparison with >1.3 million expressed sequence tag reads. We produced more than 580 million RNA-Seq reads to facilitate expression analyses. We also sequenced Setaria viridis, the ancestral wild relative of S. italica, and identified regions of differential single-nucleotide polymorphism density, distribution of transposable elements, small RNA content, chromosomal rearrangement and segregation distortion. The genus Setaria includes natural and cultivated species that demonstrate a wide capacity for adaptation. The genetic basis of this adaptation was investigated by comparing five sequenced grass genomes. We also used the diploid Setaria genome to evaluate the ongoing genome assembly of a related polyploid, switchgrass (Panicum virgatum).     

Host range of Lepidelphax pistiae (Hemiptera: Delphacidae) and its potential impact on Pistia stratiotes L. (Araceae)

Pistia stratiotes L. (Araceae) is a floating aquatic plant that has become invasive in Florida. It is primarily controlled with herbicides, but two biocontrol agents have previously been released to assist in management of this species. A new potential biocontrol agent from Argentina, Lepidelphax pistiae Remes Lenicov (Hemiptera: Delphacidae), has been evaluated comprehensively for specificity after initial host range studies done in its native range indicated that it is likely specific to P. stratiotes. Host range studies indicated that this insect is specific to P. stratiotes, with no survival or reproduction occurring on any of the 42 other plant species tested. Impact studies indicated that this insect can significantly damage P. stratiotes at medium and high population densities, which were comparable to those seen in its native range. Lepidelphax pistiae is sufficiently specific enough to warrant release and has a high probability of aiding management of P. stratiotes populations in Florida.     

Complete genome sequence of Haliscomenobacter hydrossis type strain (O)

Haliscomenobacter hydrossis van Veen et al. 1973 is the type species of the genus Haliscomenobacter, which belongs to order "Sphingobacteriales". The species is of interest because of its isolated phylogenetic location in the tree of life, especially the so far genomically uncharted part of it, and because the organism grows in a thin, hardly visible hyaline sheath. Members of the species were isolated from fresh water of lakes and from ditch water. The genome of H. hydrossis is the first completed genome sequence reported from a member of the family "Saprospiraceae". The 8,771,651 bp long genome with its three plasmids of 92 kbp, 144 kbp and 164 kbp length contains 6,848 protein-coding and 60 RNA genes, and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.     

Dissolved organic carbon in streams from artificially drained and intensively farmed watersheds in Indiana, USA

Dissolved organic carbon (DOC) in streams draining hydrologically modified and intensively farmed watersheds has not been well examined, despite the importance of these watersheds to water quality issues and the potential of agricultural soils to sequester carbon. We investigated the dynamics of DOC for 14 months during 2006 and 2007 in 6 headwater streams in a heavily agricultural and tile-drained landscape in the midwestern US. We also monitored total dissolved nitrogen (TDN) in the streams and tile drains. The concentrations of DOC in the streams and tile drains ranged from approximately 1–6 mg L^?1, while concentrations of TDN, the composition of which averaged >94% nitrate, ranged from <1 to >10 mg L^?1. Tile drains transported both DOC and TDN to the streams, but tile inputs of dissolved N were diluted by stream water, whereas DOC concentrations were generally greater in the streams than in tile drains. Filamentous algae were dense during summer base flow periods, but did not appear to contribute to the bulk DOC pool in the streams, based on diel monitoring. Short-term laboratory assays indicated that DOC in the streams was of low bioavailability, although DOC from tile drains in summer had bioavailability of 27%. We suggest that these nutrient-rich agricultural streams are well-suited for examining how increased inputs of DOC, a potential result of carbon sequestration in agricultural soils, could influence ecosystem processes.     

What free‐ranging animals do at the zoo: a study of the behavior and habitat use of opossums (Didelphis virginiana) on the grounds of the St. Louis Zoo

Studies of urban wildlife are important because they can give us insight into both how animals adapt to novel environments and how some species survive and prosper in human‐dominated landscapes. Urban zoological institutions provide an ideal setting for research on such species. We report on a study of the behavior and ecology of opossums (Didelphis virginiana) on the grounds of the St. Louis Zoological Park. We used radio tracking to investigate the movement patterns and den site use of opossums on the grounds of the zoo and compared the results to data available for opossums elsewhere. We find that urban opossums in the St. Louis zoo have smaller home ranges and move shorter distances than their rural counterparts. We suggest that increased food availability and decreased risk of predation might explain such differences, and we suspect that conditions are even more favorable to opossums in a zoological setting than in cities in general. Our findings illustrate that there is much to be learned from scientific study of the free‐living animals found on zoological park grounds. Zoo Biol 00:1–17, 2005. © 2005 Wiley‐Liss, Inc.