Global analysis of physiological responses in marine bacteria.

In this paper, we present results from studies on marine Vibrio species, in which complex adaptive responses have been investigated. The results of two-dimensional polyacrylamide gel electrophoresis serve to illustrate the usefulness of a global approach, and how it can be combined with other methodologies in order to achieve an improved understanding of the means by which bacteria adapt to alterations in environmental conditions. The overall strategies described in this paper are particularly useful for studies of bacteria for which efficient genetic tools, background genotypes and in depth physiological data are not yet available.

Synthesis of immediate upshift (Iup) proteins during recovery of marine Vibrio sp. strain S14 subjected to long-term carbon starvation.

Proteins induced during the initial phase of recovery after long-term carbon starvation in the marine Vibrio sp. strain S14 were identified by two-dimensional gel electrophoresis analysis. Nutritional upshift experiments with pulse-labeled cells were performed after addition of glucose to cells starved for 48 h. Eighteen proteins synthesized during the first 3 min after substrate addition were identified and designated immediate upshift proteins (Iup proteins). They were induced at least 10-fold compared with the rate of synthesis during starvation. Of the Iup proteins, five are not found in exponentially growing cells. Subsequent to the first 3 min of glucose addition, a complex pattern of sequential synthesis of proteins made during a transient phase as well as proteins made during 60 min of the outgrowth response was monitored. To resolve whether the Iup proteins were synthesized from stable transcripts, the initiation of transcription was inhibited by rifampin (Rif). Addition of Rif 5 min prior to glucose promoted upshift resulted in the synthesis of 12 Iup proteins. Furthermore, three Iup proteins were still induced by cells that were Rif treated 20 min prior to the upshift. These results suggest that stable but silent transcripts exist during starvation and that the translation of these mRNA species is initiated by substrate addition. This regulatory mechanism may be essential for an immediate initiation of the recovery program by the nongrowing cell.

How do non-differentiating bacteria adapt to starvation?

Non-differentiating bacteria adapt to starvation induced growth arrest by a complex turn-on/turn-off pattern of protein synthesis. This response shows distinct similarities with those of spore formation in differentiating organisms. A substantial amount of information on the non-growth biology of non-differentiating bacteria can be derived from studies on Vibrio strains. One important result is that carbon rather than nitrogen or phosphorus starvation leads to the development of a starvation and stress resistant cell in these organisms. Hence, we have attempted to characterize the carbon starvation stimulon. By the use of two-dimensional gel electrophoresis of pulse-labelled cells and transposon mutagenesis, using reporter gene constructs, the identity and function of some members of the carbon starvation stimulon have been elucidated. Moreover, regulatory genes of the starvation response have been identified with these techniques. Current studies primarily address the identity and function of these genes. The role of transcript modification and stability for both long term persistence during starvation as well as the efficient recovery of cells which occurs upon nutrient addition is also addressed. It is suggested that an understanding of the functionality of the translational machinery is essential for the understanding of these adaptive pathways. This contribution also discusses the diversity of the differentiation-like response to starvation in different bacteria and whether a general starvation induced programme exists.