William A. Cuevas

Transcriptome analysis of Crp-dependent catabolite control of gene expression in Escherichia coli.

We report here the transcriptome analyses of highly expressed genes that are subject to catabolite repression or activation mediated by the cyclic AMP receptor protein (Crp). The results reveal that many operons encoding enzymes of central carbon metabolic pathways (e.g., Krebs cycle enzymes), as well as transporters and enzymes that initiate carbon metabolism, are subject to direct Crp-mediated catabolite repression. By contrast, few enzyme-encoding genes (direct regulation) but many ribosomal protein- and tRNA-encoding genes (indirect regulation) are subject to Crp-dependent glucose activation. Additionally, Crp mediates strong indirect catabolite repression of many cytoplasmic stress response proteins, including the major chaperone proteins, five ATP-dependent protease complexes, and several cold and heat shock proteins. These results were confirmed by (i) phenotypic analyses, (ii) real-time PCR studies, (iii) reporter gene fusion assays, and (iv) previously published reports about representative genes. The results serve to define and extend our appreciation of the Crp regulon.

HARVESTER ANTS UTILIZE CUTICULAR HYDROCARBONS IN NESTMATE RECOGNITION

Cuticular hydrocarbons appear to play a role in ant nestmate recognition, but few studies have tested this hypothesis experimentally with purified hydrocarbon extracts. We exposed captive colonies of the harvester ant Pogonomyrmex barbatus to small glass blocks coated with whole cuticular lipid extracts and the purified hydrocarbon portion of extracts from nestmate and nonnestmate workers. As an estimate of agonistic behavior, we measured the proportion of ants in contact with blocks that flared their mandibles. Blocks coated with cuticular extracts from nonnestmates were contacted by more workers in one of two experiments and elicited higher levels of aggression in both experiments than blocks bearing extracts from nestmates. The cuticular hydrocarbon fraction of extracts alone was sufficient to elicit agonistic behavior toward nonnestmates. The results demonstrate that harvester ants can perceive differences in cuticular hydrocarbon composition, and can use those differences in nestmate recognition.

Functional Interactions between the Carbon and Iron Utilization Regulators, Crp and Fur, in Escherichia coli

In Escherichia coli, the ferric uptake regulator (Fur) controls expression of the iron regulon in response to iron availability while the cyclic AMP receptor protein (Crp) regulates expression of the carbon regulon in response to carbon availability. We here identify genes subject to significant changes in expression level in response to the loss of both Fur and Crp. Many iron transport genes and several carbon metabolic genes are subject to dual control, being repressed by the loss of Crp and activated by the loss of Fur. However, the sodB gene, encoding superoxide dismutase, and the aceBAK operon, encoding the glyoxalate shunt enzymes, show the opposite responses, being activated by the loss of Crp and repressed by the loss of Fur. Several other genes including the sdhA-D, sucA-D, and fumA genes, encoding key constituents of the Krebs cycle, proved to be repressed by the loss of both transcription factors. Finally, the loss of both Crp and Fur activated a heterogeneous group of genes under sigmaS control encoding, for example, the cyclopropane fatty acid synthase, Cfa, the glycogen synthesis protein, GlgS, the 30S ribosomal protein, S22, and the mechanosensitive channel protein, YggB. Many genes appeared to be regulated by the two transcription factors in an apparently additive fashion, but apparent positive or negative cooperativity characterized several putative Crp/Fur interactions. Relevant published data were evaluated, putative Crp and Fur binding sites were identified, and representative results were confirmed by real-time PCR. Molecular explanations for some, but not all, of these effects are provided.

Task-Related Differences in the Cuticular Hydrocarbon Composition of Harvester Ants, Pogonomyrmex barbatus

Colonies of the harvester ant, Pogonomyrmex barbatus, perform a variety of tasks. The behavior of an individual worker appears to depend on its recent history of brief contacts with ants of the same and other task groups. The purpose of this study was to determine whether task groups differ in cuticular hydrocarbon composition. We compared the cuticular hydrocarbon composition of ants collected under natural conditions as they performed one of three tasks: patrolling (locating food sources), foraging, or nest maintenance. Task groups differed significantly in the relative proportions of classes of hydrocarbon compounds, as well as in individual compounds. Relative to nest maintenance workers, foragers and patrollers had a higher proportion of straight-chain alkanes relative to monomethylalkanes, dimethylalkanes, and alkenes. There was no significant difference in the chain length of n-alkanes among the task groups. Foragers did not differ in hydrocarbon composition from patrollers. Colonies differed significantly from one another in hydrocarbon composition, but task groups differed in consistent ways from colony to colony, suggesting that the mechanism responsible for task-related hydrocarbon composition was the same in all colonies. P. barbatus workers switch tasks during their lifetimes, suggesting that cuticular hydrocarbon composition changes during adulthood as well. Nest maintenance workers are probably younger than foragers and patrollers and perform very little of their work outside of the nest. Task-related hydrocarbon differences detected here may be associated with worker age, and/or the abiotic characteristics (temperature, humidity, and ultraviolet light) of the interior and exterior work environments.