Crossref journal-article
Wiley
Molecular Microbiology (311)
Abstract

SummaryStrains of Pseudomonas aeruginosa causing pulmonary infections in cystic fibrosis patients have an unusual mucoid phenotype because of production of the capsule‐like exopolysaccharide, alginate. Transcriptional activation of algD, the first gene of a large alginate biosynthetic gene cluster, is associated with conversion to the alginate‐producing (Alg+) phenotype. In this study, we examined the regulation of alginate genes immediately downstream of algD. Mutants of the Alg+ strain FRD1 were constructed by gene replacement with defined Tn501 (8.2 kb) insertions in the alginate biosynthetic gene cluster, resulting in an Alg− phenotype. The Alg+ phenotype of these mutants was restored by integration of narrow‐host‐range plasmids containing DNA fragments from P. aeruginosa that reconstructed a continuous alginate gene cluster. A broad‐host‐range plasmid containing the entire alginate gene cluster except for the terminal gene, algA was unable to complement an aIG::Tn501 mutant unless algA was transcribed from a second plasmid. This indicated that any Tn501 insertion in the cluster was polar on downstream alginate genes. Northern blot hybridization experiments also showed that a transposon insertion downstream of algD adversely affected algG and algA transcription. These results provided evidence that the alginate biosynthetic gene cluster has an operonic structure and is cotranscribed from the algD promoter.

Bibliography

Chitnis, C. E., & Ohman, D. E. (1993). Genetic analysis of the alginate biosynthetic gene cluster of Pseudomonas aeruginosa shows evidence of an operonic structure. Molecular Microbiology, 8(3), 583–590. Portico.

Authors 2
  1. Chetan E. Chitnis (first)
  2. Dennis E. Ohman (additional)
References 39 Referenced 161
  1. {'key': 'e_1_2_1_2_1', 'volume-title': 'Current Protocols in Molecular Biology', 'author': 'Ausubel F.M.', 'year': '1987'} / Current Protocols in Molecular Biology by Ausubel F.M. (1987)
  2. 10.1016/0378-1119(83)90197-X
  3. 10.1093/infdis/141.2.238
  4. 10.1128/jb.171.5.2312-2317.1989
  5. 10.1128/jb.172.6.2894-2900.1990
  6. 10.1016/0378-1119(91)90290-R
  7. 10.1128/jb.161.1.249-257.1985 / J Bacteriol / Cloning of Escherichia coli and Pseudomonas aeruginosa phosphomannose isomerase genes and their expression in alginate‐negative mutants of Pseudomonas aeruginosa by Darzins A. (1985)
  8. 10.1128/jb.164.2.516-524.1985 / J Bacteriol / Clustering of mutations affecting alginic acid biosynthesis in mucoid Pseudomonas aeruginosa by Darzins A. (1985)
  9. 10.1128/jb.169.1.351-358.1987
  10. 10.1128/jb.171.3.1278-1283.1989
  11. 10.1073/pnas.70.12.3296
  12. {'key': 'e_1_2_1_13_1', 'first-page': '155', 'article-title': 'T7 early RNAs are generated by site‐specific cleavages', 'volume': '70', 'author': 'Dunne J.J.', 'year': '1973', 'journal-title': 'Proc Natl Acad Sci USA'} / Proc Natl Acad Sci USA / T7 early RNAs are generated by site‐specific cleavages by Dunne J.J. (1973)
  13. 10.1128/JB.116.2.915-924.1973
  14. 10.1073/pnas.76.4.1648 / Proc Natl Acad Sci USA / Replication of an origin‐containing derivative of plasmid RK2 dependent on a plasmid function provided in trans by Flgurski D. (1979)
  15. 10.1128/jb.170.4.1452-1460.1988
  16. 10.1128/jb.170.7.3228-3236.1988
  17. 10.1128/jb.169.4.1593-1602.1987 / J Bacteriol / Construction and characterization of Pseudomonas aeruginosa algB mutants: role of algB in high‐level production of alginate by Goldberg J.B. (1987)
  18. {'key': 'e_1_2_1_19_1', 'first-page': '302', 'article-title': 'Pseudomonas aeruginosa and cystic fibrosis: unusual bacterial adaptation and pathogenesis', 'volume': '3', 'author': 'Govan J.R.W.', 'year': '1986', 'journal-title': 'Micro Sci'} / Micro Sci / Pseudomonas aeruginosa and cystic fibrosis: unusual bacterial adaptation and pathogenesis by Govan J.R.W. (1986)
  19. 10.1073/pnas.71.3.840
  20. 10.1073/pnas.88.5.1760
  21. 10.1016/0378-1119(89)90136-4
  22. 10.1073/pnas.87.8.2887
  23. 10.1128/jb.172.5.2511-2520.1990
  24. 10.1016/0005-2787(69)90639-X
  25. {'key': 'e_1_2_1_26_1', 'volume-title': 'Molecular Cloning: A Laboratory Manual', 'author': 'Maniatis T.', 'year': '1982'} / Molecular Cloning: A Laboratory Manual by Maniatis T. (1982)
  26. 10.1128/iai.47.3.723-729.1985 / Infect Immun / Quantitation of adherence of mucoid and nonmucoid Pseudomonas aeruginosa to hamster tracheal epithelium by Marcus H. (1985)
  27. 10.1128/jb.173.16.5136-5143.1991
  28. 10.1073/pnas.70.12.3361
  29. 10.1128/IAI.33.1.142-148.1981
  30. 10.1159/000417604
  31. 10.1128/jb.162.3.1068-1074.1985 / J Bacteriol / Method for gene replacement in Pseudomonas aeruginosa used in construction of recA mutants: recA‐independent instability of alginate production by Ohman D.E. (1985)
  32. 10.1128/iai.47.1.1-4.1985 / Infect Immun / Role of Pseudomonas aeruginosa mucoid exopolysaccharide in adherence to tracheal cells by Ramphal R. (1985)
  33. 10.1016/S0021-9258(18)52212-2 / J Biol Chem / Purification and characterization of phosphomannose isomerase‐guanosine diphospho‐D‐mannose pyrophosphorylase — a bifunctional enzyme in the alginate biosynthetic pathway of Pseudomonas aeruginosa by Shinabarger D. (1991)
  34. 10.1128/iai.59.10.3596-3603.1991 / Infect Immun / Physical mapping of virulence‐associated genes in Pseudomonas aeruginosa by transverse altemating‐field electrophoresis by Shortridge V.D. (1991)
  35. {'key': 'e_1_2_1_36_1', 'first-page': '29', 'article-title': 'Alginate Inhibition of the uptake of Pseudomonas aeruginosa by macro‐phages', 'volume': '134', 'author': 'Simpson J.A.', 'year': '1988', 'journal-title': 'J Gen Microbiol'} / J Gen Microbiol / Alginate Inhibition of the uptake of Pseudomonas aeruginosa by macro‐phages by Simpson J.A. (1988)
  36. 10.1016/S0021-9258(18)65874-0 / J Biol Chem / Acetylornithinase in Escherichia coli: partial purification and some properties by Vogel H.J. (1956)
  37. {'key': 'e_1_2_1_38_1', 'first-page': '2301', 'article-title': 'Characterization of the Pseudomonas aeruginosa alginate (alg) gene region II', 'volume': '133', 'author': 'Wang S.‐K.', 'year': '1987', 'journal-title': 'J Gen Mocro biol'} / J Gen Mocro biol / Characterization of the Pseudomonas aeruginosa alginate (alg) gene region II by Wang S.‐K. (1987)
  38. 10.1128/jb.173.4.1406-1413.1991
  39. 10.1016/S0021-9258(18)92885-1
Dates
Type When
Created 18 years, 10 months ago (Oct. 27, 2006, 9:25 p.m.)
Deposited 1 year, 10 months ago (Oct. 25, 2023, 5:05 p.m.)
Indexed 1 month ago (July 31, 2025, 11:47 p.m.)
Issued 32 years, 4 months ago (May 1, 1993)
Published 32 years, 4 months ago (May 1, 1993)
Published Online 18 years, 10 months ago (Oct. 27, 2006)
Published Print 32 years, 4 months ago (May 1, 1993)
Funders 0

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@article{Chitnis_1993, title={Genetic analysis of the alginate biosynthetic gene cluster of Pseudomonas aeruginosa shows evidence of an operonic structure}, volume={8}, ISSN={1365-2958}, url={http://dx.doi.org/10.1111/j.1365-2958.1993.tb01602.x}, DOI={10.1111/j.1365-2958.1993.tb01602.x}, number={3}, journal={Molecular Microbiology}, publisher={Wiley}, author={Chitnis, Chetan E. and Ohman, Dennis E.}, year={1993}, month=may, pages={583–590} }