title: Analysis of cloned structural and regulatory genes for carbohydrate utilization in Pseudomonas aeruginosa PAO.
abstract: Five of the genes required for phosphorylative catabolism of glucose in Pseudomonas aeruginosa were ordered on two different chromosomal fragments. Analysis of a previously isolated 6.0-kb EcoRI fragment containing three structural genes showed that the genes were present on a 4.6-kb fragment in the order glucose-binding protein (gltB)-glucokinase (glk)-6-phosphogluconate dehydratase (edd). Two genes, glucose-6-phosphate dehydrogenase (zwf) and 2-keto-3-deoxy-6-phosphogluconate aldolase (eda), shown by transductional analysis to be linked to gltB and edd, were cloned on a separate 11-kb BamHI chromosomal DNA fragment and then subcloned and ordered on a 7-kb fragment. The 6.0-kb EcoRI fragment had been shown to complement a regulatory mutation, hexR, which caused noninducibility of four glucose catabolic enzymes. In this study, hexR was mapped coincident with edd. A second regulatory function, hexC, was cloned within a 0.6-kb fragment contiguous to the edd gene but containing none of the structural genes. The phenotypic effect of the hexC locus, when present on a multicopy plasmid, was elevated expression of glucokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydratase, and 2-keto-3-deoxy-6-phosphogluconate aldolase activities in the absence of inducer.
authors.affiliation: Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond 23298.
authors.forename: L
authors.initials: L
authors.lastname: Temple
authors.forename: S M
authors.initials: SM
authors.lastname: Cuskey
authors.forename: R E
authors.initials: RE
authors.lastname: Perkins
authors.forename: R C
authors.initials: RC
authors.lastname: Bass
authors.forename: N M
authors.initials: NM
authors.lastname: Morales
authors.forename: G E
authors.initials: GE
authors.lastname: Christie
authors.forename: R H
authors.initials: RH
authors.lastname: Olsen
authors.forename: P V
authors.initials: PV
authors.lastname: Phibbs
mesh_terms: D050260:Carbohydrate Metabolism; D002678:Chimera; D002876:Chromosomes, Bacterial; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005798:Genes, Bacterial; D005809:Genes, Regulator; D005838:Genotype; D005947:Glucose / Q000378:metabolism*; D008954:Models, Biological; D010957:Plasmids; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping
chemical_list: D004269:DNA, Bacterial; D005947:Glucose