Bridging factorial and gradient concepts of resource co‐limitation: towards a general framework applied to consumers

Organism growth can be limited either by a single resource or by multiple resources simultaneously (co‐limitation). Efforts to characterise co‐limitation have generated two influential approaches. One approach uses limitation scenarios of factorial growth assays to distinguish specific types of co‐limitation; the other uses growth responses spanned over a continuous, multi‐dimensional resource space to characterise different types of response surfaces. Both approaches have been useful in investigating particular aspects of co‐limitation, but a synthesis is needed to stimulate development of this recent research area. We address this gap by integrating the two approaches, thereby presenting a more general framework of co‐limitation. We found that various factorial (co‐)limitation scenarios can emerge in different response surface types based on continuous availabilities of essential or substitutable resources. We tested our conceptual co‐limitation framework on data sets of published and unpublished studies examining the limitation of two herbivorous consumers in a two‐dimensional resource space. The experimental data corroborate the predictions, suggesting a general applicability of our co‐limitation framework to generalist consumers and potentially also to other organisms. The presented framework might give insight into mechanisms that underlie co‐limitation responses and thus can be a seminal starting point for evaluating co‐limitation patterns in experiments and nature.     

Cell-free synthesis, membrane integration, and glycosylation of pro-sucrase-isomaltase

Cell-free translation of total RNA from rabbit intestinal mucosa in a rabbit reticulocyte lysate, after immunoprecipitation with antibodies directed against sucrase-isomaltase, yielded a polypeptide of 200 kDa, which was identified as pro-sucrase-isomaltase. Addition of dog pancreatic microsomal vesicles to the translation system resulted in the appearance of an additional 220-kDa polypeptide. The 220-kDa polypeptide was associated with the membranes in a way that made it inaccessible to proteolysis; this protection was abolished by lytic detergent concentrations, indicating that the polypeptide was segregated into the microsomal vesicle. The 220-kDa polypeptide was glycosylated as evidenced by it being bound to concanavalin A-Sepharose and eluted with alpha-methyl-D-mannopyranoside. The increase in apparent molecular mass (approximately 20 kDa) of the primary translation product upon translocation was due to the addition of carbohydrate; treatment of the 220-kDa polypeptide with endo-beta-N-acetylglucosaminidase H increased its electrophoretic mobility to that of the 200-kDa polypeptide which was obtained in the absence of membranes. Partial N-terminal amino acid sequence of a translation product labeled with [3H]Leu in the absence of membranes revealed that Leu was incorporated into identical positions as in the final (pro)-sucrase-isomaltase, thus indicating the lack of a transient signal peptide.     

Photodynamic action and bacterial protein synthesis

Summary Photodynamic treatment of bacterial cells with thiopyronin and psoralen inhibits protein synthesis. Transfer RNA isolated from thiopyronin- and psoralen-treated cells is inactive in cell-free protein synthesis. Both the sensitizers inactivate transfer RNA to about 60 to 70%. Ribosomes isolated from thiopyronin-sensitized cells lose 45% of their activity, whereas under the influence of psoralen only 20% inactivation is observed. The enzyme fraction is not damaged in psoralen-treated bacteria, but thiopyronin inactivates this fraction as well. In vivo experiments indicate that psoralen does not react with the protein components of the cell. The mechanism of these inactivations is discussed.     

From sextant to GPS: Twenty‐five years of mapping the genome with ChIP

Since its inception, ChIP technology has evolved immensely. Technological advances have improved its specificity and sensitivity, its scale has expanded to a genome‐wide level, and its relative ease of use has made it a virtually ubiquitous tool. This year marks the 25th anniversary of the development of ChIP. In honor of this milestone, we briefly revisit its history, offer a review of recent articles employing ChIP on a genome‐wide scale, and lay out our views for the future of ChIP. J. Cell. Biochem. 107: 6–10, 2009. © 2009 Wiley‐Liss, Inc.     

Detecting antigens by quantitative immuno-PCR

The quantitative immuno-PCR (qIPCR) technology combines the advantages of flexible and robust immunoassays with the exponential signal amplification power of PCR. The qIPCR allows one to detect antigens using specific antibodies labeled with double-stranded DNA. The label is used for signal generation by quantitative PCR. Because of the efficiency of nucleic acid amplification, qIPCR typically leads to a 10- to 1,000-fold increase in sensitivity compared to an analogous enzyme-amplified immunoassay. A standard protocol of a qIPCR assay to detect human interleukin 6 (IL-6) using a sandwich immunoassay combined with real-time PCR readout is described here. The protocol includes initial immobilization of the antigen, and coupling of this antigen with antibody-DNA conjugates is then carried out by (a) the stepwise assembly of biotinylated antibody, streptavidin and biotinylated DNA, (b) the use of a biotinylated antibody and an anti-biotin-DNA conjugate or (c) the employment of an anti-IL-6 antibody-DNA conjugate. Following the assembly of signal-generating immunocomplexes, real-time PCR is used to amplify and record the signal. Depending on the coupling strategy, the qIPCR assays require 4-7 h with only about 3 h hands-on-time. The use of qIPCR assays enables the detection of rare biomarkers in complex biological samples that are poorly accessible by conventional immunoassays. Therefore, qIPCR offers novel opportunities for the biomedical analysis of, for instance, neurodegenerative diseases and viral infections as well as new tools for the development of novel pharmaceuticals.     

Characterization of Bacillus subtilis mutants with carbon source-independent glutamate biosynthesis

Bacillus subtilis synthesizes glutamate from 2-oxoglutarate and glutamine using the glutamate synthase, encoded by the gltAB operon. Glutamate degradation involves the catabolic glutamate dehydrogenase (GDH) RocG. Expression of both gltAB and rocG is controlled by the carbon and nitrogen sources. In the absence of glucose or other well-metabolizable carbon sources, B. subtilis is unable to grow unless provided with external glutamate. In this work, we isolated mutations that suppressed this growth defect of B. subtilis on minimal media (sgd mutants). All mutations enabled the cells to express the gltAB operon even in the absence of glucose. The mutations were all identified in the rocG gene suggesting that the catabolic GDH is essential for controlling gltAB expression in response to the availability of sugars.     

Vasopressin, oxytocin, and social odor recognition

Central vasopressin and oxytocin, and their homologues, modulate a multitude of social behaviors in a variety of animal taxa. All social behavior requires some level of social (re)cognition, and these neuropeptides exert powerful effects on an animal's ability to recognize and appropriately respond to a conspecific. Social cognition for many mammals, including rodents, begins at the main and accessory olfactory systems. We recently identified vasopressin expressing neurons in the main and accessory olfactory bulb and in the anterior olfactory nucleus, a region of olfactory cortex that transmits and processes information in the main olfactory system. We review this and other work demonstrating that both vasopressin and oxytocin modulate conspecific social recognition at the level of the olfactory system. We also outline recent work on the somato-dendritic release of vasopressin and oxytocin, and propose a model by which the somato-dendritic priming of these neuropeptides in main olfactory regions may facilitate the formation of short-term social odor memories. This article is part of a Special Issue entitled Oxytocin, Vasopressin, and Social Behavior.