Distinct conformations of the kinesin Unc104 neck regulate a monomer to dimer motor transition

Caenhorhabditis elegans Unc104 kinesin transports synaptic vesicles at rapid velocities. Unc104 is primarily monomeric in solution, but recent motility studies suggest that it may dimerize when concentrated on membranes. Using cryo-electron microscopy, we observe two conformations of microtubule-bound Unc104: a monomeric state in which the two neck helices form an intramolecular, parallel coiled coil; and a dimeric state in which the neck helices form an intermolecular coiled coil. The intramolecular folded conformation is abolished by deletion of a flexible hinge separating the neck helices, indicating that it acts as a spacer to accommodate the parallel coiled-coil configuration. The neck hinge deletion mutation does not alter motor velocity in vitro but produces a severe uncoordinated phenotype in transgenic C. elegans, suggesting that the folded conformation plays an important role in motor regulation. We suggest that the Unc104 neck regulates motility by switching from a self-folded, repressed state to a dimerized conformation that can support fast processive movement.     

Lead compounds discovered from libraries: part 2

Many lead compounds with the potential to progress to viable drug candidates have been identified from libraries during the past two years. There are two key strategies most often employed to find leads from libraries: first, high-throughput biological screening of corporate compound collections; and second, synthesis and screening of project-directed libraries (i.e. target-based libraries). Numerous success stories, including the discovery of several clinical candidates, testify to the utility of chemical library collections as proven sources of new leads for drug development.     

Managing N inputs and the effect on N losses following excretion in open-dirt feedlots in Nebraska

Nutrition will play an important role in meeting the environmental challenges of beef cattle feedlots. Nutritionists are continually refining protein requirements, and have recently adopted a new metabolizable protein (MP) system to more efficiently use nitrogen (N) and allow more accurate diet formulation. Protein requirements vary by animal age and weight during the finishing period. Our hypothesis was that formulating diets with the MP system would decrease N inputs and lead to decreased excretion and losses. Comparing industry average diets (13.5% crude protein) to phase-fed diets formulated to not exceed MP requirements decreased N inputs by 10 to 20% for calves and yearlings without affecting average daily gain. Decreasing inputs led to a concomitant decrease in N excretion (12 to 21%) and losses (15 to 33%) in open-dirt feedlot pens. N losses are variable with time of year, with averages of 60 to 70% of excreted N lost during the summer months and 40% lost during the November to May feeding periods. Protein requirements are being refined continually as more research data are collected. However, formulation to meet protein requirements, but not exceed them, is an important nutritional management option for feedlots to become sustainable.     

Fatty acids enhanced tubermycin production by Pseudomonas strain 2HS

A new microbial isolate, Pseudomonas 2HS, produced trace amounts of a greenish-yellow pigment when grown aerobically in a 1% yeast extract medium at 30 degrees C and shaken at 250 rpm for 5 days. In contrast, cells produced more greenish-yellow pigment (2.16 mg/15 ml culture) when grown in the presence of 0.5% 12-hydroxyoctadecanoic acid (w/v). The greenish-yellow pigment was identified as phenazine-1-carboxylic acid (tubermycin B), and the Pseudomonas 2HS was identified as P. aeruginosa 2HS. This is the first report that 12-hydroxyoctadecanoic, ricinoleic and other fatty acids can enhance the production of phenazine-1-carboxylic acid by a Pseudomonas species.     

Multiple, non-allelic, intein-coding sequences in eukaryotic RNA polymerase genes

Background  

Inteins are self-splicing protein elements. They are translated as inserts within host proteins that excise themselves and ligate the flanking portions of the host protein (exteins) with a peptide bond. They are encoded as in-frame insertions within the genes for the host proteins. Inteins are found in all three domains of life and in viruses, but have a very sporadic distribution. Only a small number of intein coding sequences have been identified in eukaryotic nuclear genes, and all of these are from ascomycete or basidiomycete fungi.     

Method for recovering sulfolipid from plant lipid extracts

Sulfoquinovosyl diglyceride from plant tissues is recovered in sizeable quantities, free from phospholipids and galactolipids.