PMID:10352233

Citation	Vazquez, A, Moreno, S, Guzmán, J, Alvarado, A and Espín, G (1999) Transcriptional organization of the Azotobacter vinelandii algGXLVIFA genes: characterization of algF mutants. Gene 232:217-22
Abstract	Azotobacter vinelandii forms desiccation-resistant cysts which contain a high proportion of the exopolysaccharide alginate in their envelope. We have previously shown that the A. vinelandii alginate biosynthetic genes algA and algL are transcribed from a promoter located somewhere upstream of algL. In this study we sequenced the A. vinelandii algX, algL, algV, algI and algF genes located between algG and algA. We carried out primer extension analysis of the algG, algX and algL genes and detected transcription start sites upstream algG but not upstream algX or algL, implying that algG and algX form part of the previously identified algL-A operon. A promoter upstream algA was also detected; however, transcription of algA exclusively from this promoter is not sufficient for the AlgA levels required for alginate production. An algF mutant (AJ34) was constructed by insertion of the Omega-tetracycline cassette in the non-polar orientation. As expected, AJ34 produced unacetylated alginate. Viability of 35day old cysts formed by strain AJ34, but not of those formed by the wild type, was reduced, indicating that acetylation of alginate plays a role in cyst resistance to desiccation.
Links	PubMed
Keywords	Alginates/metabolism; Azotobacter vinelandii/genetics; Azotobacter vinelandii/physiology; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Base Sequence; Carbohydrate Epimerases/chemistry; Carbohydrate Epimerases/genetics; Cloning, Molecular; Codon, Initiator/isolation & purification; Genes, Bacterial; Mannose-6-Phosphate Isomerase/chemistry; Mannose-6-Phosphate Isomerase/genetics; Molecular Sequence Data; Multienzyme Complexes/chemistry; Multienzyme Complexes/genetics; Multigene Family; Mutation; Nucleotidyltransferases/chemistry; Nucleotidyltransferases/genetics; Operon; Plasmids/chemical synthesis; RNA, Messenger/isolation & purification; Spores, Bacterial/physiology; Transcription, Genetic
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
AZOVI:ALGF		GO:0051979: alginic acid acetylation	ECO:0000315:		P		Table 2: Strain AJ34 carrying the algF::ΩTc insertion in the non-polar orientation produced unacetylated alginate. Strain AJ21 carrying the algF::ΩTc mutation in the polar orientation did not produce alginate. Plasmid pRSA55 carrying the A. vinelandii algL-A region was transferred by conjugation to strain AJ34. This plasmid restored wild-type mucoid phenotype and alginate acetylation to AJ34 mutant.	complete CACAO 5004
AZOVI:ALGF	involved_in	GO:0051979: alginic acid acetylation	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

References

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