Abstract
Phospholipid synthesis has been reported to be subject to stringent control in Escherichia coli. We present evidence that demonstrates a strict correlation between guanosine tetraphosphate accumulation and inhibition of phospholipid synthesis. In vivo experiments designed to examine the pattern of phospholipid labeling with 32P-inorganic phosphate and 32P-sn-glycerol-3-phosphate suggest that regulation must occur at the glycerol-3-phosphate acyltransferase step. Assay of phospholipid synthesis by cell-free extracts and semipurified preparations revealed that guanosine tetraphosphate inhibits at least two enzymes specific for the biosynthetic pathway, sn-glycerol-3-phosphate acyltransferase as well as sn-glycerol-3-phosphate phosphatidyl transferase. These findings provide a biochemical basis for the stringent control of lipid synthesis as well as regulation of steady-state levels of phospholipid in growing cells.
Full text
PDF![355](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/c8a6c5cfe576/jbacter00345-0371.png)
![356](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/792f19ea41d8/jbacter00345-0372.png)
![357](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/4f71752010c4/jbacter00345-0373.png)
![358](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/01c25779f217/jbacter00345-0374.png)
![359](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/ebaf07010eb5/jbacter00345-0375.png)
![360](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/810f3ffb235f/jbacter00345-0376.png)
![361](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/9b2f16e146c9/jbacter00345-0377.png)
![362](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/27c37cdd62e7/jbacter00345-0378.png)
![363](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/8f838258c346/jbacter00345-0379.png)
![364](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/995d711b3f4e/jbacter00345-0380.png)
![365](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/10b1f8e180df/jbacter00345-0381.png)
![366](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/429f/246430/338b3da35ea6/jbacter00345-0382.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ailhaud G. P., Vagelos P. R. Palmityl-acyl carrier protein as acyl donor for complex lipid biosynthesis in Escherichia coli. J Biol Chem. 1966 Aug 25;241(16):3866–3869. [PubMed] [Google Scholar]
- Ames G. F. Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism. J Bacteriol. 1968 Mar;95(3):833–843. doi: 10.1128/jb.95.3.833-843.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Atherly A. G. Temperature-sensitive relaxed Phenotype in a stringent strain of Escherichia coli. J Bacteriol. 1973 Jan;113(1):178–182. doi: 10.1128/jb.113.1.178-182.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
- Ballesta J. P., Schaechter M. Dependence of the rate of synthesis of phosphatidylethanolamine and phosphatidylglycerol on the rate of growth of Escherichia coli. J Bacteriol. 1972 Apr;110(1):452–453. doi: 10.1128/jb.110.1.452-453.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cashel M., Gallant J. Two compounds implicated in the function of the RC gene of Escherichia coli. Nature. 1969 Mar 1;221(5183):838–841. doi: 10.1038/221838a0. [DOI] [PubMed] [Google Scholar]
- Cashel M., Kalbacher B. The control of ribonucleic acid synthesis in Escherichia coli. V. Characterization of a nucleotide associated with the stringent response. J Biol Chem. 1970 May 10;245(9):2309–2318. [PubMed] [Google Scholar]
- Cashel M., Lazzarini R. A., Kalbacher B. An improved method for thin-layer chromatography of nucleotide mixtures containing 32P-labelled orthophosphate. J Chromatogr. 1969 Mar 11;40(1):103–109. doi: 10.1016/s0021-9673(01)96624-5. [DOI] [PubMed] [Google Scholar]
- Cashel M. The control of ribonucleic acid synthesis in Escherichia coli. IV. Relevance of unusual phosphorylated compounds from amino acid-starved stringent strains. J Biol Chem. 1969 Jun 25;244(12):3133–3141. [PubMed] [Google Scholar]
- Chang Y. Y., Kennedy E. P. Biosynthesis of phosphatidyl glycerophosphate in Escherichia coli. J Lipid Res. 1967 Sep;8(5):447–455. [PubMed] [Google Scholar]
- Cronan J. E., Jr, Ray T. K., Vagelos P. R. Selection and characterization of an E. coli mutant defective in membrane lipid biosynthesis. Proc Natl Acad Sci U S A. 1970 Mar;65(3):737–744. doi: 10.1073/pnas.65.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donini P. Amino acid control over deoxyribonucleic acid synthesis in Escherichia coli infected with T-even bacteriophage. J Bacteriol. 1970 Jun;102(3):616–627. doi: 10.1128/jb.102.3.616-627.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edlin G., Donini P. Synthesis of guanosine 5'-diphosphate, 2'-(or 3'-) diphosphate and related nucleotides in a variety of physiological conditions. J Biol Chem. 1971 Jul 10;246(13):4371–4373. [PubMed] [Google Scholar]
- Gallant J., Harada B. The control of ribonucleic acid synthesis in Escherichia coli. 3. The functional relationship between purine ribonucleoside triphosphate pool sizes and the rate of ribonucleic acid accumulation. J Biol Chem. 1969 Jun 25;244(12):3125–3132. [PubMed] [Google Scholar]
- Gallant J., Irr J., Cashel M. The mechanism of amino acid control of guanylate and adenylate biosynthesis. J Biol Chem. 1971 Sep 25;246(18):5812–5816. [PubMed] [Google Scholar]
- Gallant J., Margason G., Finch B. On the turnover of ppGpp in Escherichia coli. J Biol Chem. 1972 Oct 10;247(19):6055–6058. [PubMed] [Google Scholar]
- Glaser M., Bayer W. H., Bell R. M., Vagelos P. R. Regulation of macromolecular biosynthesis in a mutant of Escherichia coli defective in membrane phospholipid biosynthesis. Proc Natl Acad Sci U S A. 1973 Feb;70(2):385–389. doi: 10.1073/pnas.70.2.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glynn I. M., Chappell J. B. A simple method for the preparation of 32-P-labelled adenosine triphosphate of high specific activity. Biochem J. 1964 Jan;90(1):147–149. doi: 10.1042/bj0900147. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Golden N. G., Powell G. L. Stringent and relaxed control of phospholipid metabolism in Escherichia coli. J Biol Chem. 1972 Oct 25;247(20):6651–6658. [PubMed] [Google Scholar]
- Goldfine H. Use of a filter-paper disk assay in the measurement of lipid biosynthesis. J Lipid Res. 1966 Jan;7(1):146–149. [PubMed] [Google Scholar]
- Harshman R. B., Yamazaki H. Formation of ppGpp in a relaxed and stringent strain of Escherichia coli during diauxie lag. Biochemistry. 1971 Oct 12;10(21):3980–3982. doi: 10.1021/bi00797a027. [DOI] [PubMed] [Google Scholar]
- Haseltine W. A., Block R., Gilbert W., Weber K. MSI and MSII made on ribosome in idling step of protein synthesis. Nature. 1972 Aug 18;238(5364):381–384. doi: 10.1038/238381a0. [DOI] [PubMed] [Google Scholar]
- Haseltine W. A., Block R. Synthesis of guanosine tetra- and pentaphosphate requires the presence of a codon-specific, uncharged transfer ribonucleic acid in the acceptor site of ribosomes. Proc Natl Acad Sci U S A. 1973 May;70(5):1564–1568. doi: 10.1073/pnas.70.5.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hochstadt-Ozer J., Cashel M. The regulation of purine utilization in bacteria. V. Inhibition of purine phosphoribosyltransferase activities and purine uptake in isolated membrane vesicles by guanosine tetraphosphate. J Biol Chem. 1972 Nov 10;247(21):7067–7072. [PubMed] [Google Scholar]
- KANFER J., KENNEDY E. P. METABOLISM AND FUNCTION OF BACTERIAL LIPIDS. II. BIOSYNTHESIS OF PHOSPHOLIPIDS IN ESCHERICHIA COLI. J Biol Chem. 1964 Jun;239:1720–1726. [PubMed] [Google Scholar]
- Kito M., Lubin M., Pizer L. I. A mutant of Escherichia coli defective in phosphatidic acid synthesis. Biochem Biophys Res Commun. 1969 Feb 21;34(4):454–458. doi: 10.1016/0006-291x(69)90403-3. [DOI] [PubMed] [Google Scholar]
- Kito M., Pizer L. I. Phosphatidic acid synthesis in Escherichia coli. J Bacteriol. 1969 Mar;97(3):1321–1327. doi: 10.1128/jb.97.3.1321-1327.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kito M., Pizer L. I. Purification and regulatory properties of the biosynthetic L-glycerol 3-phosphate dehydrogenase from Escherichia coli. J Biol Chem. 1969 Jun 25;244(12):3316–3323. [PubMed] [Google Scholar]
- Lazzarini R. A., Cashel M., Gallant J. On the regulation of guanosine tetraphosphate levels in stringent and relaxed strains of Escherichia coli. J Biol Chem. 1971 Jul 25;246(14):4381–4385. [PubMed] [Google Scholar]
- Lowry O. H., Carter J., Ward J. B., Glaser L. The effect of carbon and nitrogen sources on the level of metabolic intermediates in Escherichia coli. J Biol Chem. 1971 Nov;246(21):6511–6521. [PubMed] [Google Scholar]
- Mavis R. D., Vagelos P. R. The effect of phospholipid fatty acid composition in membranous enzymes in Escherichia coli. J Biol Chem. 1972 Feb 10;247(3):652–659. [PubMed] [Google Scholar]
- Pedersen F. S., Lund E., Kjeldgaard N. O. Codon specific, tRNA dependent in vitro synthesis of ppGpp and pppGpp. Nat New Biol. 1973 May 2;243(122):13–15. [PubMed] [Google Scholar]
- Pizer L. I., Merlie J. P. Effect of serine hydroxamate on phospholipid synthesis in Escherichia coli. J Bacteriol. 1973 Jun;114(3):980–987. doi: 10.1128/jb.114.3.980-987.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rabbani E., Srinivasan P. R. Role of the translocation factor G in the regulation of ribonucleic acid synthesis. J Bacteriol. 1973 Mar;113(3):1177–1183. doi: 10.1128/jb.113.3.1177-1183.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sokawa J., Sokawa Y., Kaziro Y. Stringent control in Escherichia coli. Nat New Biol. 1972 Dec 20;240(103):242–245. doi: 10.1038/newbio240242a0. [DOI] [PubMed] [Google Scholar]
- Sokawa Y., Nakao E., Kaziro Y. On the nature of the control by RC gene in e. coli: amino acid-dependent control of lipid synthesis. Biochem Biophys Res Commun. 1968 Oct 10;33(1):108–112. doi: 10.1016/0006-291x(68)90263-5. [DOI] [PubMed] [Google Scholar]
- Sy J., Lipmann F. Identification of the synthesis of guanosine tetraphosphate (MS I) as insertion of a pyrophosphoryl group into the 3'-position in guanosine 5'-diphosphate. Proc Natl Acad Sci U S A. 1973 Feb;70(2):306–309. doi: 10.1073/pnas.70.2.306. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tosa T., Pizer L. I. Effect of serine hydroxamate on the growth of Escherichia coli. J Bacteriol. 1971 Jun;106(3):966–971. doi: 10.1128/jb.106.3.966-971.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winslow R. M. A consequence of the rel gene during a glucose to lactate downshift in Escherichia coli. The rates of ribonucleic acid synthesis. J Biol Chem. 1971 Aug 10;246(15):4872–4877. [PubMed] [Google Scholar]