J. Bacteriol., Mar 1996, 1401-1411, Vol 178, No. 5
Copyright © 1996, American Society for Microbiology

Functional analysis of a relA/spoT gene homolog from Streptococcus equisimilis

U Mechold, M Cashel, K Steiner, D Gentry and H Malke
Institute for Molecular Biology, Jena University, Germany.

We examined the functional attributes of a gene encountered by sequencing the streptokinase gene region of Streptococcus equisimilis H46A. This gene, originally called rel, here termed relS. equisimilis, is homologous to two related Escherichia coli genes, spoT and relA, that function in the metabolism of guanosine 5',3'-polyphosphates [(p)ppGpp]. Studies with a variety of E. coli mutants led us to deduce that the highly expressed rel S. equisimilis gene encodes a strong (p)ppGppase and a weaker (p)ppGpp synthetic activity, much like the spoT gene, with a net effect favoring degradation and no complementation of the absence of the relA gene. We verified that the Rel S. equisimilis protein, purified from an E. coli relA spoT double mutant, catalyzed a manganese-activated (p)ppGpp 3'- pyrophosphohydrolase reaction similar to that of the SpoT enzyme. This Rel S. equisimilis protein preparation also weakly catalyzed a ribosome- independent synthesis of (p)ppGpp by an ATP to GTP 3'- pyrophosphoryltransferase reaction when degradation was restricted by the absence of manganese ions. An analogous activity has been deduced for the SpoT protein from genetic evidence. In addition, the Rel S. equisimilis protein displays immunological cross-reactivity with polyclonal antibodies specific for SpoT but not for RelA. Despite assignment of rel S. equisimilis gene function in E. coli as being similar to that of the native spoT gene, disruptions of rel S. equisimilis in S. equisimilis abolish the parental (p)ppGpp accumulation response to amino acid starvation in a manner expected for relA mutants rather than spoT mutants.

This article has been cited by other articles:

• Gomez-Escribano, J. P., Martin, J. F., Hesketh, A., Bibb, M. J., Liras, P. (2008). Streptomyces clavuligerus relA-null mutants overproduce clavulanic acid and cephamycin C: negative regulation of secondary metabolism by (p)ppGpp. Microbiology 154: 744-755 [Abstract] [Full Text]  
• Nascimento, M. M., Lemos, J. A., Abranches, J., Lin, V. K., Burne, R. A. (2008). Role of RelA of Streptococcus mutans in Global Control of Gene Expression. J. Bacteriol. 190: 28-36 [Abstract] [Full Text]  
• Burne, R. A., Bessen, D. E., Broadbent, J. R., Claverys, J.-P. (2007). The Seventh International Conference on the Genetics of Streptococci, Lactococci, and Enterococci. J. Bacteriol. 189: 1209-1218 [Full Text]  
• Jain, V., Saleem-Batcha, R., China, A., Chatterji, D. (2006). Molecular dissection of the mycobacterial stringent response protein Rel.. Protein Sci. 15: 1449-1464 [Abstract] [Full Text]  
• Kim, S., Watanabe, K., Suzuki, H., Watarai, M. (2005). Roles of Brucella abortus SpoT in morphological differentiation and intramacrophagic replication. Microbiology 151: 1607-1617 [Abstract] [Full Text]  
• Proudfoot, M., Kuznetsova, E., Brown, G., Rao, N. N., Kitagawa, M., Mori, H., Savchenko, A., Yakunin, A. F. (2004). General Enzymatic Screens Identify Three New Nucleotidases in Escherichia coli: BIOCHEMICAL CHARACTERIZATION OF SurE, YfbR, AND YjjG. J. Biol. Chem. 279: 54687-54694 [Abstract] [Full Text]  
• Yakunin, A. F., Proudfoot, M., Kuznetsova, E., Savchenko, A., Brown, G., Arrowsmith, C. H., Edwards, A. M. (2004). The HD Domain of the Escherichia coli tRNA Nucleotidyltransferase Has 2',3'-Cyclic Phosphodiesterase, 2'-Nucleotidase, and Phosphatase Activities. J. Biol. Chem. 279: 36819-36827 [Abstract] [Full Text]  
• Lemos, J. A. C., Brown, T. A. Jr., Burne, R. A. (2004). Effects of RelA on Key Virulence Properties of Planktonic and Biofilm Populations of Streptococcus mutans. Infect. Immun. 72: 1431-1440 [Abstract] [Full Text]  
• Concepcion, M. B., Nelson, D. R. (2003). Expression of spoT in Borrelia burgdorferi during Serum Starvation. J. Bacteriol. 185: 444-452 [Abstract] [Full Text]  
• Ajdic', D., McShan, W. M., McLaughlin, R. E., Savic', G., Chang, J., Carson, M. B., Primeaux, C., Tian, R., Kenton, S., Jia, H., Lin, S., Qian, Y., Li, S., Zhu, H., Najar, F., Lai, H., White, J., Roe, B. A., Ferretti, J. J. (2002). Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc. Natl. Acad. Sci. USA 99: 14434-14439 [Abstract] [Full Text]  
• Potrykus, K., Santos, S., Lemonnier, M., Diaz-Orejas, R., Wegrzyn, G. (2002). Differential effects of Kid toxin on two modes of replication of lambdoid plasmids suggest that this toxin acts before, but not after, the assembly of the replication complex. Microbiology 148: 2489-2495 [Abstract] [Full Text]  
• Bugrysheva, J., Dobrikova, E. Y., Godfrey, H. P., Sartakova, M. L., Cabello, F. C. (2002). Modulation of Borrelia burgdorferi Stringent Response and Gene Expression during Extracellular Growth with Tick Cells. Infect. Immun. 70: 3061-3067 [Abstract] [Full Text]  
• Mechold, U., Murphy, H., Brown, L., Cashel, M. (2002). Intramolecular Regulation of the Opposing (p)ppGpp Catalytic Activities of RelSeq, the Rel/Spo Enzyme from Streptococcus equisimilis. J. Bacteriol. 184: 2878-2888 [Abstract] [Full Text]  
• Taylor, C. M., Beresford, M., Epton, H. A. S., Sigee, D. C., Shama, G., Andrew, P. W., Roberts, I. S. (2002). Listeria monocytogenes relA and hpt Mutants Are Impaired in Surface-Attached Growth and Virulence. J. Bacteriol. 184: 621-628 [Abstract] [Full Text]  
• Steiner, K., Malke, H. (2001). relA-Independent Amino Acid Starvation Response Network of Streptococcus pyogenes. J. Bacteriol. 183: 7354-7364 [Abstract] [Full Text]  
• Sun, J., Hesketh, A., Bibb, M. (2001). Functional Analysis of relA and rshA, Two relA/spoT Homologues of Streptomyces coelicolor A3(2). J. Bacteriol. 183: 3488-3498 [Abstract] [Full Text]  
• Wehmeier, L., Brockmann-Gretza, O., Pisabarro, A., Tauch, A., Pühler, A., Martin, J. F., Kalinowski, J. (2001). A Corynebacterium glutamicum mutant with a defined deletion within the rplK gene is impaired in (p)ppGpp accumulation upon amino acid starvation. Microbiology 147: 691-700 [Abstract] [Full Text]  
• Primm, T. P., Andersen, S. J., Mizrahi, V., Avarbock, D., Rubin, H., Barry, C. E. III (2000). The Stringent Response of Mycobacterium tuberculosis Is Required for Long-Term Survival. J. Bacteriol. 182: 4889-4898 [Abstract] [Full Text]  
• Gentry, D., Li, T., Rosenberg, M., McDevitt, D. (2000). The rel Gene Is Essential for In Vitro Growth of Staphylococcus aureus. J. Bacteriol. 182: 4995-4997 [Abstract] [Full Text]  
• Ojha, A. K., Mukherjee, T. K., Chatterji, D. (2000). High Intracellular Level of Guanosine Tetraphosphate in Mycobacterium smegmatis Changes the Morphology of the Bacterium. Infect. Immun. 68: 4084-4091 [Abstract] [Full Text]  
• Venkateswaran, A., McFarlan, S. C., Ghosal, D., Minton, K. W., Vasilenko, A., Makarova, K., Wackett, L. P., Daly, M. J. (2000). Physiologic Determinants of Radiation Resistance in Deinococcus radiodurans. Appl. Environ. Microbiol. 66: 2620-2626 [Abstract] [Full Text]  
• Sen, K., Hayashi, J.-i., Kuramitsu, H. K. (2000). Characterization of the relA Gene of Porphyromonas gingivalis. J. Bacteriol. 182: 3302-3304 [Abstract] [Full Text]  
• Martínez-Costa, O. H., Fernández-Moreno, M. A., Malpartida, F. (1998). The relA/spoT-Homologous Gene in Streptomyces coelicolor Encodes Both Ribosome-Dependent (p)ppGppSynthesizing and -Degrading Activities. J. Bacteriol. 180: 4123-4132 [Abstract] [Full Text]