THE EFFECTS OF 3,4-DIHYDROXYBUTYL 1-PHOSPHONATE ON ESCHERICHIA COLI. A CORRELATION OF 3,4-DIHYDROXYBUTYL 1-PHOSPHONATE RESISTANCE WITH A DEFECT IN CARDIOLIPIN SYNTHESIS.
Item
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Title
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THE EFFECTS OF 3,4-DIHYDROXYBUTYL 1-PHOSPHONATE ON ESCHERICHIA COLI. A CORRELATION OF 3,4-DIHYDROXYBUTYL 1-PHOSPHONATE RESISTANCE WITH A DEFECT IN CARDIOLIPIN SYNTHESIS.
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Identifier
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AAI8501141
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identifier
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8501141
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Creator
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HWANG, YU-WEN.
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Contributor
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Burton E. Tropp
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Date
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1984
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Microbiology
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Abstract
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Escherichia coli treated for 1 h with 100 uM rac-3,4-dihydroxybutyl 1-phosphonate (DBP), a glycerol 3-phosphate analog, die when stored at 5(DEGREES)C, whereas the viability of untreated cells is relatively unaffected. This observation formed the basis of a selection procedure that was used to isolate mutants that are partially resistant to DBP. One such mutant, strain 6204, is constitutive for DBP transport, exhibits a particularly high degree of cold resistance, has the same doubling time as the parent and is similar to the parent strain in terms of incorporation of DBP into the lipid fraction. Glycerol 3-phosphate and phosphatidylglycerol phosphate synthetases obtained from 6204 and its parent were identical in terms of DBP recognition. The parent strain is killed when incubated in the presence of a combination of 70 uM rac-DBP and 0.25% deoxycholate, whereas strain 6204 continues to grow, albeit more slowly, in the presences of this combination. Strain 6204 can be distinguished from the parent strain on agar plates (low phosphate minimal medium with glucuronate as the sole carbon source) containing 15 uM rac-DBP.;The insertion of Tn10 near to the 6204 mutation has facilitated genetic manipulations. All the phenotypic effects attributed to strain 6204 appear to be due to a single mutation. Genetic analysis indicates that Tn10, inserted near to the gene responsible for DBP resistance, maps in the vicinity of min 27. Three factor crosses reveal a gene order of hemA-Dbp('r) -Tn10 (zch) -trp. The only gene for phosphoglyceride metabolism, known to map in this region, is the gene associated with cardiolipin synthetase, cls. Genetic results suggest that the mutation responsible for DBP resistance maps in or very near to cls. analysis of the lipids isolated from untreated strain 6204 (and from each of the transductants prepared by Plvir mediated transfer of DBP-resistance to wild type strains) reveal that cardiolipin (CL) synthesis is defective. These results strongly suggest that the mutation responsible for DBP resistance has its primary effect on CL synthesis. To further test this hypothesis, strains with an authentic cls mutation were constructed and examined for resistance to DBP. These strains had growth properties that were identical with those of strain 6204. . . . (Author's abstract exceeds stipulated maximum length. Discontinued here with permission of author.) UMI.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
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Program
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Biochemistry
