Search citation statements
Paper Sections
Citation Types
Year Published
Publication Types
Relationship
Authors
Journals
Fatty acid degradation was investigated in Caulobacter crescentus, a bacterium that exhibits membranemediated differentiation events. Two strains of C. crescentus were shown to utilize oleic acid as sole carbon source. Five enzymes of the fatty acid 13-oxidation pathway, acyl-coenzyme A (CoA) synthase, crotonase, thiolase, 3-hydroxyacyl-CoA dehydrogenase, and acyl-CoA dehydrogenase, were identified. The activities of these enzymes were significantly higher in C. crescentus than the fully induced levels observed in Escherichia coli. Growth in glucose or glucose plus oleic acid decreased fatty acid uptake and lowered the specific activity of the enzymes involved in 1-oxidation by 2-to 3-fold, in contrast to the 50-fold glucose repression found in E. coli. The mild glucose repression of the acyl-CoA synthase was reversed by exogenous dibutyryl cyclic AMP. Acyl-CoA synthase activity was shown to be the same in oleic acid-grown cells and in cells grown in the presence of succinate, a carbon source not affected by catabolite repression. Thus, fatty acid degradation by the 13-oxidation pathway is constitutive in C. crescentus and is only mildly affected by growth in the presence of glucose. TnS insertion mutants unable to form colonies when oleic acid was the sole carbon source were isolated. However, these mutants efficiently transported fatty acids and had 1-oxidation enzyme levels comparable with that of the wild type. Our inability to obtain fatty acid degradation mutants after a wide search, coupled with the high constitutive levels of the 1-oxidation enzymes, suggest that fatty acid turnover, as has proven to be the case in fatty acid biosynthesis, might play an essential role in membrane biogenesis and cell cycle events in C. crescentus.The cell cycle of the gram-negative bacterium Caulobacter crescentus exhibits a series of unicellular differentiation events. Several of these events involve changes at the cell surface and include the biogenesis of a polar flagellum, chemosensory receptors, phage receptors, and pili (23). The synthesis of many of the membrane proteins involved in these structures and functions is temporally regulated. Upon synthesis these proteins are localized to specific portions of the cell and segregate to only one daughter cell upon division (23). Thus, some type of spatial control mechanism must contribute to their positioning in the cell membrane. To help define the role that the cell membrane might play in these processes, we investigated the composition and metabolism of the membrane phospholipids.The negatively charged phospholipids, phosphatidylglycerol and cardiolipin, are the major components, and neither phosphatidylethanolamine nor phosphatidylserine can be detected in C. crescentus membranes (6). The enzymatic pathway of phospholipid synthesis in C. crescentus is the same as the comparable portions of the pathway in Escherichia coli (5). The fatty acid components of the phospholipids consist of both saturated and unsaturated 14-, 16-, and 18-carbon fatty acids (4, 16), and these ...
Fatty acid degradation was investigated in Caulobacter crescentus, a bacterium that exhibits membranemediated differentiation events. Two strains of C. crescentus were shown to utilize oleic acid as sole carbon source. Five enzymes of the fatty acid 13-oxidation pathway, acyl-coenzyme A (CoA) synthase, crotonase, thiolase, 3-hydroxyacyl-CoA dehydrogenase, and acyl-CoA dehydrogenase, were identified. The activities of these enzymes were significantly higher in C. crescentus than the fully induced levels observed in Escherichia coli. Growth in glucose or glucose plus oleic acid decreased fatty acid uptake and lowered the specific activity of the enzymes involved in 1-oxidation by 2-to 3-fold, in contrast to the 50-fold glucose repression found in E. coli. The mild glucose repression of the acyl-CoA synthase was reversed by exogenous dibutyryl cyclic AMP. Acyl-CoA synthase activity was shown to be the same in oleic acid-grown cells and in cells grown in the presence of succinate, a carbon source not affected by catabolite repression. Thus, fatty acid degradation by the 13-oxidation pathway is constitutive in C. crescentus and is only mildly affected by growth in the presence of glucose. TnS insertion mutants unable to form colonies when oleic acid was the sole carbon source were isolated. However, these mutants efficiently transported fatty acids and had 1-oxidation enzyme levels comparable with that of the wild type. Our inability to obtain fatty acid degradation mutants after a wide search, coupled with the high constitutive levels of the 1-oxidation enzymes, suggest that fatty acid turnover, as has proven to be the case in fatty acid biosynthesis, might play an essential role in membrane biogenesis and cell cycle events in C. crescentus.The cell cycle of the gram-negative bacterium Caulobacter crescentus exhibits a series of unicellular differentiation events. Several of these events involve changes at the cell surface and include the biogenesis of a polar flagellum, chemosensory receptors, phage receptors, and pili (23). The synthesis of many of the membrane proteins involved in these structures and functions is temporally regulated. Upon synthesis these proteins are localized to specific portions of the cell and segregate to only one daughter cell upon division (23). Thus, some type of spatial control mechanism must contribute to their positioning in the cell membrane. To help define the role that the cell membrane might play in these processes, we investigated the composition and metabolism of the membrane phospholipids.The negatively charged phospholipids, phosphatidylglycerol and cardiolipin, are the major components, and neither phosphatidylethanolamine nor phosphatidylserine can be detected in C. crescentus membranes (6). The enzymatic pathway of phospholipid synthesis in C. crescentus is the same as the comparable portions of the pathway in Escherichia coli (5). The fatty acid components of the phospholipids consist of both saturated and unsaturated 14-, 16-, and 18-carbon fatty acids (4, 16), and these ...
We have identified mutations in three pleiotropic genes, pleA, pkC, and pleD, that are required for differentiation in Caulobacter crescentus. pleA and pleC mutants were isolated in an extensive screen for strains defective in both motility and adsorption of polar bacteriophage +CbK; using temperature-sensitive alleles, we determined the time at which the two genes act. pleA was required for a short period at 0.7 of the swarmer cell cycle for flagellum biosynthesis, whereas pleC was required during an overlapping period from 0.6 to 0.95 of the cell cycle to activate flagellum rotation as well as to enable loss of the flagellum and stalk formation by swarmer cells after division. The third pleiotropic gene, pleD, is described here for the first time. A pleD mutation was identified as a bypass suppressor of a temperature-sensitive pleC allele. Strains containing this mutation were highly motile, did not shed the flagellum or form stalks, and retained motility throughout the cell cycle. Since pleD was required to turn off motility and was a bypass suppressor of pleC, we conclude that it acts after the pleA and pleC gene functions in the cell cycle. No mutants defective in both flagellum biosynthesis and stalk formation were identified. Consequently, we propose that the steps required for formation of swarmer cells and subsequent development into stalked cells are organized into at least two developmental pathways: a pleA-dependent sequence of events, responsible for flagellum biosynthesis in predivisional cells, and a pleC-pleD-dependent sequence, responsible for flagellum activation in predivisional cells and loss of motility and stalk formation in progeny swarmer cells.
Several Caulobacter crescentus mutants with lesions in phospholipid biosynthesis have DNA replication phenotypes. A C. crescentus mutant deficient in glycerol 3-phosphate dehydrogenase activity (gpsA) blocks phospholipid synthesis, ceases DNA replication, and loses viability in the absence of a glycerol phosphate supplement. To investigate the interaction between membrane synthesis and DNA replication during a single cell cycle, we moved the gpsA mutation into a synchronizable, but otherwise wild-type, strain. The first effect of withholding supplement was the cessation of synthesis of phosphatidylglycerol, a major component of the C. crescentus membrane. In the absence of glycerol 3-phosphate, DNA replication was initiated in the stalked cell at the correct time in the cell cycle and at the correct site on the chromosome. However, after replication proceeded bidirectionally for a short time, DNA synthesis dropped to a low level. The cell cycle blocked at a distinct middivision stalked cell, and this was followed by cell death. The "glycerol-less" death of the gpsA mutant could be prevented if the cells were treated with novobiocin to prevent the initiation of DNA replication. Our observations suggest that the processivity of C. crescentus replication requires concomitant phospholipid synthesis and that cell death results from incomplete replication of the chromosome.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.