Nucleotide sequence and expression of the Pseudomonas aeruginosa algF gene controlling acetylation of alginate.
Colonization of the cystic fibrosis lung by Pseudomonas aeruginosa is greatly facilitated by the production of an exopolysaccharide called alginate. In this study we determined the nucleotide sequence of an alginate modification gene, algF, which controls the addition of acetyl groups to alginate. Expression of algF using a T7 promoter-expression system showed that algF codes for a 24.5 kDa polypeptide (predicted size 22,832 Da) that is processed to 19.5 kDa. The N-terminus of the processed polypeptide matched the predicted amino acid sequence of AlgF starting at Asp-29. An algF mutant failed to produce alginate owing to a polar effect on the downstream algA gene. Although the algA gene, provided in trans, restored synthesis of alginate, the alginate was non-acetylated. We show that a plasmid containing both the algF and algA gene complements the alginate acetylation defect of the algF mutant strain.
Shinabarger
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Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago 60612.
May
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Boyd
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Ghosh
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Chakrabarty
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AM
D000107:Acetylation; D000464:Alginates / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003550:Cystic Fibrosis / Q000382:microbiology; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004926:Escherichia coli; D015870:Gene Expression; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D008168:Lung / Q000382:microbiology; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping
D000464:Alginates; D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D006603:Hexuronic Acids; C089910:algF protein, Pseudomonas aeruginosa; D020723:Glucuronic Acid