Metabolic context and possible physiological themes of sigma(54)-dependent genes in Escherichia coli

Abstract

Sigma(54) has several features that distinguish it from other sigma factors in Escherichia coli: it is not homologous to other sigma subunits, sigma(54)-dependent expression absolutely requires an activator, and the activator binding sites can be far from the transcription start site. A rationale for these properties has not been readily apparent, in part because of an inability to assign a common physiological function for sigma(54)-dependent genes. Surveys of sigma(54)-dependent genes from a variety of organisms suggest that the products of these genes are often involved in nitrogen assimilation; however, many are not. Such broad surveys inevitably remove the sigma(54)-dependent genes from a potentially coherent metabolic context. To address this concern, we consider the function and metabolic context of sigma(54)-dependent genes primarily from a single organism, Escherichia coli, in which a reasonably complete list of sigma(54)-dependent genes has been identified by computer analysis combined with a DNA microarray analysis of nitrogen limitation-induced genes. E. coli appears to have approximately 30 sigma(54)-dependent operons, and about half are involved in nitrogen assimilation and metabolism. A possible physiological relationship between sigma(54)-dependent genes may be based on the fact that nitrogen assimilation consumes energy and intermediates of central metabolism. The products of the sigma(54)-dependent genes that are not involved in nitrogen metabolism may prevent depletion of metabolites and energy resources in certain environments or partially neutralize adverse conditions. Such a relationship may limit the number of physiological themes of sigma(54)-dependent genes within a single organism and may partially account for the unique features of sigma(54) and sigma(54)-dependent gene expression.

FIG. 1

ς⁵⁴-dependent genes in E. coli. The known ς⁵⁴-dependent operons and their transcripts are shown in the context of neighboring genes. The open and solid boxes indicate counterclockwise and clockwise transcription, respectively. If the gene has been assigned a function, this is indicated by the gene name underneath. Boxes without gene names indicate that the gene has an unknown function. The sizes of the genes and their intergenic regions are to scale.

FIG. 2

Regulatory regions of the characterized ς⁵⁴-dependent genes. The following features are shown for each of the ς⁵⁴-dependent promoters: binding sites for activators, DNA bending proteins (when required), and RNA polymerase. Solid boxes indicate that the binding site has been demonstrated. A hatched box indicates a confirmed binding site, but binding is weak. An open box signifies a proposed binding site. The relative location and orientation of the nearest upstream gene are also shown. All diagrams are to the same scale, except for the hyp operon.

FIG. 3

Distribution of scores with properly and improperly oriented sites. The open bars indicate sites that are oriented toward the 3′ end of a gene; the solid bars indicate sites that are oriented toward the 5′ end.

FIG. 4

Pathways of ammonia assimilation. GDH, glutamate dehydrogenase.

FIG. 5

Regulation of GS activity and the Ntr response. The pathways are shown for conditions of nitrogen excess (high glutamine) (top) and nitrogen limitation (low glutamine) with partial GlnK uridylylation (bottom). The open arrow in the bottom panel is meant to indicate that only partial uridylylation occurs. It is assumed that partial uridylylation occurs either during nitrogen limitation or during the transition to steady-state nitrogen-limited growth. The T-like symbol indicates an inhibition.

FIG. 6

Binding sites for regulatory proteins at the hutUH promoter.

FIG. 7

Metabolic relationships between ornithine, arginine, putrescine, and GABA. A thick black arrow indicates that nitrogen limitation induces the enzyme indicated. A reaction catalyzed by two (or more) enzymes is indicated by two arrows. The genes that specify the enzymes are shown when they are known. A dashed arrow indicates that the gene has yet to be identified.