Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Nov;174(21):7063–7067. doi: 10.1128/jb.174.21.7063-7067.1992

Resolution of chromosomes III and VI of Aspergillus nidulans by pulsed-field gel electrophoresis shows that the penicillin biosynthetic pathway genes pcbAB, pcbC, and penDE are clustered on chromosome VI (3.0 megabases).

E Montenegro 1, F Fierro 1, F J Fernandez 1, S Gutiérrez 1, J F Martín 1
PMCID: PMC207390  PMID: 1400258

Abstract

An improved electrophoretic molecular karyotype of Aspergillus nidulans ATCC 28901 has been obtained by contour-clamped electric field gel electrophoresis, which separates seven chromosomal bands and allows resolution of chromosomes III and VI. The three genes of the penicillin biosynthetic pathway, pcbAB, pcbC, and penDE, encoding alpha-aminoadipyl-cysteinyl-valine synthetase, isopenicillin N synthase, and isopenicillin N acyltransferase, respectively, are clustered together on a chromosome of 3.0 Mg, corresponding to linkage group VI, whereas the argB gene was located on a chromosome of 3.4 Mb, corresponding to linkage group III. Three other strains of A. nidulans contained a modified chromosome III of about 3.1 Mb that overlaps with chromosome VI, forming a doublet. Resolution of chromosomes III and VI in strain ATCC 28901 allowed unequivocal mapping of the penicillin gene cluster on chromosome VI of A. nidulans.

Full text

PDF
7066

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barredo J. L., Díez B., Alvarez E., Martín J. F. Large amplification of a 35-kb DNA fragment carrying two penicillin biosynthetic genes in high penicillin producing strains of Penicillium chrysogenum. Curr Genet. 1989 Dec;16(5-6):453–459. doi: 10.1007/BF00340725. [DOI] [PubMed] [Google Scholar]
  2. Barredo J. L., van Solingen P., Díez B., Alvarez E., Cantoral J. M., Kattevilder A., Smaal E. B., Groenen M. A., Veenstra A. E., Martín J. F. Cloning and characterization of the acyl-coenzyme A: 6-aminopenicillanic-acid-acyltransferase gene of Penicillium chrysogenum. Gene. 1989 Nov 30;83(2):291–300. doi: 10.1016/0378-1119(89)90115-7. [DOI] [PubMed] [Google Scholar]
  3. Brody H., Carbon J. Electrophoretic karyotype of Aspergillus nidulans. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6260–6263. doi: 10.1073/pnas.86.16.6260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chu G., Vollrath D., Davis R. W. Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science. 1986 Dec 19;234(4783):1582–1585. doi: 10.1126/science.3538420. [DOI] [PubMed] [Google Scholar]
  5. Díez B., Barredo J. L., Alvarez E., Cantoral J. M., van Solingen P., Groenen M. A., Veenstra A. E., Martín J. F. Two genes involved in penicillin biosynthesis are linked in a 5.1 kb SalI fragment in the genome of Penicillium chrysogenum. Mol Gen Genet. 1989 Sep;218(3):572–576. doi: 10.1007/BF00332426. [DOI] [PubMed] [Google Scholar]
  6. Díez B., Gutiérrez S., Barredo J. L., van Solingen P., van der Voort L. H., Martín J. F. The cluster of penicillin biosynthetic genes. Identification and characterization of the pcbAB gene encoding the alpha-aminoadipyl-cysteinyl-valine synthetase and linkage to the pcbC and penDE genes. J Biol Chem. 1990 Sep 25;265(27):16358–16365. [PubMed] [Google Scholar]
  7. Gutiérrez S., Díez B., Montenegro E., Martín J. F. Characterization of the Cephalosporium acremonium pcbAB gene encoding alpha-aminoadipyl-cysteinyl-valine synthetase, a large multidomain peptide synthetase: linkage to the pcbC gene as a cluster of early cephalosporin biosynthetic genes and evidence of multiple functional domains. J Bacteriol. 1991 Apr;173(7):2354–2365. doi: 10.1128/jb.173.7.2354-2365.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. KAFER E. An 8-chromosome map of Aspergillus nidulans. Adv Genet. 1958;9:105–145. [PubMed] [Google Scholar]
  9. MacCabe A. P., Riach M. B., Unkles S. E., Kinghorn J. R. The Aspergillus nidulans npeA locus consists of three contiguous genes required for penicillin biosynthesis. EMBO J. 1990 Jan;9(1):279–287. doi: 10.1002/j.1460-2075.1990.tb08106.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MacCabe A. P., van Liempt H., Palissa H., Unkles S. E., Riach M. B., Pfeifer E., von Döhren H., Kinghorn J. R. Delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase from Aspergillus nidulans. Molecular characterization of the acvA gene encoding the first enzyme of the penicillin biosynthetic pathway. J Biol Chem. 1991 Jul 5;266(19):12646–12654. [PubMed] [Google Scholar]
  11. Makins J. F., Allsop A., Holt G. Intergeneric cosynthesis of penicillin by strains of Penicillium chrysogenum, P. chrysogenum/notatum and Aspergillus nidulans. J Gen Microbiol. 1981 Feb;122(2):339–343. doi: 10.1099/00221287-122-2-339. [DOI] [PubMed] [Google Scholar]
  12. Makins J. F., Holt G., Macdonald K. D. The genetic location of three mutations impairing penicillin production in Aspergillus nidulans. J Gen Microbiol. 1983 Oct;129(10):3027–3033. doi: 10.1099/00221287-129-10-3027. [DOI] [PubMed] [Google Scholar]
  13. Martín M. F., Liras P. Organization and expression of genes involved in the biosynthesis of antibiotics and other secondary metabolites. Annu Rev Microbiol. 1989;43:173–206. doi: 10.1146/annurev.mi.43.100189.001133. [DOI] [PubMed] [Google Scholar]
  14. Montenegro E., Barredo J. L., Gutiérrez S., Díez B., Alvarez E., Martín J. F. Cloning, characterization of the acyl-CoA:6-amino penicillanic acid acyltransferase gene of Aspergillus nidulans and linkage to the isopenicillin N synthase gene. Mol Gen Genet. 1990 May;221(3):322–330. doi: 10.1007/BF00259395. [DOI] [PubMed] [Google Scholar]
  15. Mortimer R. K., Schild D. Genetic map of Saccharomyces cerevisiae, edition 9. Microbiol Rev. 1985 Sep;49(3):181–213. doi: 10.1128/mr.49.3.181-213.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Orbach M. J., Vollrath D., Davis R. W., Yanofsky C. An electrophoretic karyotype of Neurospora crassa. Mol Cell Biol. 1988 Apr;8(4):1469–1473. doi: 10.1128/mcb.8.4.1469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ramón D., Carramolino L., Patiño C., Sánchez F., Peñalva M. A. Cloning and characterization of the isopenicillin N synthetase gene mediating the formation of the beta-lactam ring in Aspergillus nidulans. Gene. 1987;57(2-3):171–181. doi: 10.1016/0378-1119(87)90120-x. [DOI] [PubMed] [Google Scholar]
  18. Schwartz D. C., Cantor C. R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. doi: 10.1016/0092-8674(84)90301-5. [DOI] [PubMed] [Google Scholar]
  19. Smith D. J., Burnham M. K., Bull J. H., Hodgson J. E., Ward J. M., Browne P., Brown J., Barton B., Earl A. J., Turner G. Beta-lactam antibiotic biosynthetic genes have been conserved in clusters in prokaryotes and eukaryotes. EMBO J. 1990 Mar;9(3):741–747. doi: 10.1002/j.1460-2075.1990.tb08168.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Snell R. G., Wilkins R. J. Separation of chromosomal DNA molecules from C.albicans by pulsed field gel electrophoresis. Nucleic Acids Res. 1986 Jun 11;14(11):4401–4406. doi: 10.1093/nar/14.11.4401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Timberlake W. E. Low repetitive DNA content in Aspergillus nidulans. Science. 1978 Dec 1;202(4371):973–975. doi: 10.1126/science.362530. [DOI] [PubMed] [Google Scholar]
  22. Tobin M. B., Fleming M. D., Skatrud P. L., Miller J. R. Molecular characterization of the acyl-coenzyme A:isopenicillin N acyltransferase gene (penDE) from Penicillium chrysogenum and Aspergillus nidulans and activity of recombinant enzyme in Escherichia coli. J Bacteriol. 1990 Oct;172(10):5908–5914. doi: 10.1128/jb.172.10.5908-5914.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Van der Ploeg L. H., Schwartz D. C., Cantor C. R., Borst P. Antigenic variation in Trypanosoma brucei analyzed by electrophoretic separation of chromosome-sized DNA molecules. Cell. 1984 May;37(1):77–84. doi: 10.1016/0092-8674(84)90302-7. [DOI] [PubMed] [Google Scholar]
  24. Vollrath D., Davis R. W. Resolution of DNA molecules greater than 5 megabases by contour-clamped homogeneous electric fields. Nucleic Acids Res. 1987 Oct 12;15(19):7865–7876. doi: 10.1093/nar/15.19.7865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weigel B. J., Burgett S. G., Chen V. J., Skatrud P. L., Frolik C. A., Queener S. W., Ingolia T. D. Cloning and expression in Escherichia coli of isopenicillin N synthetase genes from Streptomyces lipmanii and Aspergillus nidulans. J Bacteriol. 1988 Sep;170(9):3817–3826. doi: 10.1128/jb.170.9.3817-3826.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yelton M. M., Hamer J. E., Timberlake W. E. Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1470–1474. doi: 10.1073/pnas.81.5.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. van Gorcom R. F., Punt P. J., Pouwels P. H., van den Hondel C. A. A system for the analysis of expression signals in Aspergillus. Gene. 1986;48(2-3):211–217. doi: 10.1016/0378-1119(86)90079-x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES