title: Inactivation of sll1556 in Synechocystis strain PCC 6803 impairs isoprenoid biosynthesis from pentose phosphate cycle substrates in vitro.
abstract: In cyanobacteria many compounds, including chlorophylls, carotenoids, and hopanoids, are synthesized from the isoprenoid precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate. Isoprenoid biosynthesis in extracts of the cyanobacterium Synechocystis strain PCC 6803 grown under photosynthetic conditions, stimulated by pentose phosphate cycle substrates, does not appear to require methylerythritol phosphate pathway intermediates. The sll1556 gene, distantly related to type 2 IPP isomerase genes, was disrupted by insertion of a Kanr cassette. The mutant was fully viable under photosynthetic conditions although impaired in the utilization of pentose phosphate cycle substrates. Compared to the parental strain the Deltasll1556 mutant (i) is deficient in isoprenoid biosynthesis in vitro with substrates including glyceraldehyde-3-phosphate, fructose-6-phosphate, and glucose-6-phosphate; (ii) has smaller cells (diameter ca. 13% less); (iii) has fewer thylakoids (ca. 30% less); and (iv) has a more extensive fibrous outer wall layer. Isoprenoid biosynthesis is restored with pentose phosphate cycle substrates plus the recombinant Sll1556 protein in the Deltasll1556 supernatant fraction. IPP isomerase activity could not be demonstrated for the purified Sll1556 protein under our in vitro conditions. The reduction of thylakoid area and the effect on outer wall layer components are consistent with an impairment of isoprenoid biosynthesis in the mutant, possibly via hopanoid biosynthesis. Our findings are consistent with an alternate metabolic shunt for biosynthesis of isoprenoids.
authors.affiliation: Department of Cell Biology and Molecular Genetics, Microbiology Building, Campus Dr., University of Maryland, College Park, MD 20742, USA.
authors.forename: Kelly
authors.initials: K
authors.lastname: Poliquin
authors.forename: Yuri V
authors.initials: YV
authors.lastname: Ershov
authors.forename: Francis X
authors.initials: FX
authors.lastname: Cunningham
authors.forename: Tinsay T
authors.initials: TT
authors.lastname: Woreta
authors.forename: R Raymond
authors.initials: RR
authors.lastname: Gantt
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Gantt
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D002338:Carotenoids / Q000096:biosynthesis; D002473:Cell Wall / Q000648:ultrastructure; D002734:Chlorophyll / Q000096:biosynthesis; D000458:Cyanobacteria / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D005636:Fructosephosphates / Q000378:metabolism; D020868:Gene Silencing; D005798:Genes, Bacterial / Q000235:genetics / Q000502:physiology*; D019298:Glucose-6-Phosphate / Q000378:metabolism; D005986:Glyceraldehyde 3-Phosphate / Q000378:metabolism; D045782:Hemiterpenes / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009943:Organophosphorus Compounds / Q000378:metabolism; D010427:Pentose Phosphate Pathway / Q000502:physiology*; D013729:Terpenes / Q000378:metabolism*; D020524:Thylakoids / Q000648:ultrastructure
chemical_list: D001426:Bacterial Proteins; D005636:Fructosephosphates; D045782:Hemiterpenes; D009943:Organophosphorus Compounds; D013729:Terpenes; D002734:Chlorophyll; D005986:Glyceraldehyde 3-Phosphate; C004809:isopentenyl pyrophosphate; C043060:3,3-dimethylallyl pyrophosphate; D002338:Carotenoids; D019298:Glucose-6-Phosphate; C027618:fructose-6-phosphate