Abstract
The stringent control response, which involves a rapid accumulation of ppGpp, is triggered if the marine Vibrio sp. strain S14 is subjected to carbon and energy starvation. By means of high-resolution two-dimensional gel electrophoresis analysis, we addressed the role of the major ppGpp-synthesizing enzyme (RelA) in the regulation of the carbon starvation response of Vibrio sp. strain S14. The finding that a large number of the carbon starvation-induced proteins were underexpressed in the Vibrio sp. S14 relA mutant strain after the onset of glucose starvation suggests that a rapid accumulation of ppGpp is required for induction of many of the carbon starvation-induced proteins. However, it was also found that a majority of the carbon starvation-induced proteins were significantly less induced if the stringent control response was provoked by amino acid starvation. We therefore also addressed the notion that a carbon starvation-specific signal transduction pathway, complementary to the stringent control, may exist in Vibrio sp. strain S14. It was found that a majority of the proteins that were underexpressed in the relA mutant strain were also underexpressed in the Vibrio sp. S14 spoT mutant strain (csrS1). Interestingly, a large proportion of these underexpressed proteins were found to belong to a group of proteins that are not, or significantly less, induced by starvation conditions that do not promote starvation survival. On the basis of these observations and the finding that the csrS1 strain survives poorly but accumulates ppGpp in a fashion similar to the wild type during carbon and energy source starvation, the gene product of the csrS gene is suggested to be responsible for the mediation of a signal which is complementary to ppGpp and essential for the successful development of the starvation- and stress-resistant cell. This conclusion was also supported by experiments in which changes in phenotypic characteristics known to be induced during carbon starvation were studied. The starvation induction of the high-affinity glucose uptake system was found to be dependent on the csrS gene but not relA, and the synthesis of carbon starvation-specific periplasmic space proteins was dependent, at different times of starvation, on both the relA and the csrS gene products.
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- Albertson N. H., Nyström T., Kjelleberg S. Exoprotease Activity of Two Marine Bacteria during Starvation. Appl Environ Microbiol. 1990 Jan;56(1):218–223. doi: 10.1128/aem.56.1.218-223.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Albertson N. H., Nyström T., Kjelleberg S. Starvation-induced modulations in binding protein-dependent glucose transport by the marine Vibrio sp. S14. FEMS Microbiol Lett. 1990 Jul;58(2):205–209. doi: 10.1111/j.1574-6968.1990.tb13979.x. [DOI] [PubMed] [Google Scholar]
- Button D. K. Biochemical basis for whole-cell uptake kinetics: specific affinity, oligotrophic capacity, and the meaning of the michaelis constant. Appl Environ Microbiol. 1991 Jul;57(7):2033–2038. doi: 10.1128/aem.57.7.2033-2038.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chaloner-Larsson G., Yamazaki H. Effects of the spoT and relA mutation on the synthesis and accumulation of ppGpp and RNA during glucose starvation. Can J Biochem. 1978 Apr;56(4):264–272. doi: 10.1139/o78-041. [DOI] [PubMed] [Google Scholar]
- Flärdh K., Axberg T., Albertson N. H., Kjelleberg S. Stringent control during carbon starvation of marine Vibrio sp. strain S14: molecular cloning, nucleotide sequence, and deletion of the relA gene. J Bacteriol. 1994 Oct;176(19):5949–5957. doi: 10.1128/jb.176.19.5949-5957.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garrels J. I. The QUEST system for quantitative analysis of two-dimensional gels. J Biol Chem. 1989 Mar 25;264(9):5269–5282. [PubMed] [Google Scholar]
- Geesey G. G., Morita R. Y. Capture of arginine at low concentrations by a marine psychrophilic bacterium. Appl Environ Microbiol. 1979 Dec;38(6):1092–1097. doi: 10.1128/aem.38.6.1092-1097.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Givskov M., Eberl L., Molin S. Responses to nutrient starvation in Pseudomonas putida KT2442: two-dimensional electrophoretic analysis of starvation- and stress-induced proteins. J Bacteriol. 1994 Aug;176(16):4816–4824. doi: 10.1128/jb.176.16.4816-4824.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Groat R. G., Schultz J. E., Zychlinsky E., Bockman A., Matin A. Starvation proteins in Escherichia coli: kinetics of synthesis and role in starvation survival. J Bacteriol. 1986 Nov;168(2):486–493. doi: 10.1128/jb.168.2.486-493.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hernandez V. J., Bremer H. Escherichia coli ppGpp synthetase II activity requires spoT. J Biol Chem. 1991 Mar 25;266(9):5991–5999. [PubMed] [Google Scholar]
- Holmquist L., Nelson D. R., Kjelleberg S. The DnaK homologue of the marine Vibrio sp. strain S14 binds to the unprocessed form of a carbon starvation-specific periplasmic protein. Mol Microbiol. 1994 Mar;11(5):861–868. doi: 10.1111/j.1365-2958.1994.tb00364.x. [DOI] [PubMed] [Google Scholar]
- Molin S., Von Meyenburg K., Maaloe O., Hansen M. T., Pato M. L. Control of ribosome synthesis in Escherichia coli: analysis of an energy source shift-down. J Bacteriol. 1977 Jul;131(1):7–17. doi: 10.1128/jb.131.1.7-17.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morton D. S., Oliver J. D. Induction of Carbon Starvation-Induced Proteins in Vibrio vulnificus. Appl Environ Microbiol. 1994 Oct;60(10):3653–3659. doi: 10.1128/aem.60.10.3653-3659.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nyström T., Olsson R. M., Kjelleberg S. Survival, stress resistance, and alterations in protein expression in the marine vibrio sp. strain S14 during starvation for different individual nutrients. Appl Environ Microbiol. 1992 Jan;58(1):55–65. doi: 10.1128/aem.58.1.55-65.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nystöm T. Role of guanosine tetraphosphate in gene expression and the survival of glucose or seryl-tRNA starved cells of Escherichia coli K12. Mol Gen Genet. 1994 Nov 1;245(3):355–362. doi: 10.1007/BF00290116. [DOI] [PubMed] [Google Scholar]
- Ostling J., Flärdh K., Kjelleberg S. Isolation of a carbon starvation regulatory mutant in a marine Vibrio strain. J Bacteriol. 1995 Dec;177(23):6978–6982. doi: 10.1128/jb.177.23.6978-6982.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spector M. P., Cubitt C. L. Starvation-inducible loci of Salmonella typhimurium: regulation and roles in starvation-survival. Mol Microbiol. 1992 Jun;6(11):1467–1476. doi: 10.1111/j.1365-2958.1992.tb00867.x. [DOI] [PubMed] [Google Scholar]
- Xiao H., Kalman M., Ikehara K., Zemel S., Glaser G., Cashel M. Residual guanosine 3',5'-bispyrophosphate synthetic activity of relA null mutants can be eliminated by spoT null mutations. J Biol Chem. 1991 Mar 25;266(9):5980–5990. [PubMed] [Google Scholar]