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. 1997 Nov;65(11):4718–4724. doi: 10.1128/iai.65.11.4718-4724.1997

Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus.

J A Calera 1, S Paris 1, M Monod 1, A J Hamilton 1, J P Debeaupuis 1, M Diaquin 1, R López-Medrano 1, F Leal 1, J P Latgé 1
PMCID: PMC175677  PMID: 9353056

Abstract

Aspergillus fumigatus possesses two catalases (described as fast and slow on the basis of their electrophoretic mobility). The slow catalase has been recognized as a diagnostic antigen for aspergillosis in immunocompetent patients. The antigenic catalase has been purified. The enzyme is a tetrameric protein composed of 90-kDa subunits. The corresponding cat1 gene was cloned, and sequencing data show that the cat1 gene codes for a 728-amino-acid polypeptide. A recombinant protein expressed in Pichia pastoris is enzymatically active and has biochemical and antigenic properties that are similar to those of the wild-type catalase. Molecular experiments reveal that CAT1 contains a signal peptide and a propeptide of 15 and 12 amino acid residues, respectively. cat1-disrupted mutants that were unable to produce the slow catalase were as sensitive to H2O2 and polymorphonuclear cells as the wild-type strain. In addition, there was no difference in pathogenicity between the cat1 mutant and its parental cat1+ strain in a murine model of aspergillosis.

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Selected References

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  1. Arruda L. K., Platts-Mills T. A., Fox J. W., Chapman M. D. Aspergillus fumigatus allergen I, a major IgE-binding protein, is a member of the mitogillin family of cytotoxins. J Exp Med. 1990 Nov 1;172(5):1529–1532. doi: 10.1084/jem.172.5.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beauvais A., Monod M., Debeaupuis J. P., Diaquin M., Kobayashi H., Latgé J. P. Biochemical and antigenic characterization of a new dipeptidyl-peptidase isolated from Aspergillus fumigatus. J Biol Chem. 1997 Mar 7;272(10):6238–6244. doi: 10.1074/jbc.272.10.6238. [DOI] [PubMed] [Google Scholar]
  3. Carrez D., Janssens W., Degrave P., van den Hondel C. A., Kinghorn J. R., Fiers W., Contreras R. Heterologous gene expression by filamentous fungi: secretion of human interleukin-6 by Aspergillus nidulans. Gene. 1990 Oct 15;94(2):147–154. doi: 10.1016/0378-1119(90)90381-z. [DOI] [PubMed] [Google Scholar]
  4. Fita I., Rossmann M. G. The active center of catalase. J Mol Biol. 1985 Sep 5;185(1):21–37. doi: 10.1016/0022-2836(85)90180-9. [DOI] [PubMed] [Google Scholar]
  5. Fowler T., Rey M. W., Vähä-Vahe P., Power S. D., Berka R. M. The catR gene encoding a catalase from Aspergillus niger: primary structure and elevated expression through increased gene copy number and use of a strong promoter. Mol Microbiol. 1993 Sep;9(5):989–998. doi: 10.1111/j.1365-2958.1993.tb01228.x. [DOI] [PubMed] [Google Scholar]
  6. Fuller R. S., Brake A., Thorner J. Yeast prohormone processing enzyme (KEX2 gene product) is a Ca2+-dependent serine protease. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1434–1438. doi: 10.1073/pnas.86.5.1434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gavel Y., von Heijne G. Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering. Protein Eng. 1990 Apr;3(5):433–442. doi: 10.1093/protein/3.5.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Girardin H., Latgé J. P., Srikantha T., Morrow B., Soll D. R. Development of DNA probes for fingerprinting Aspergillus fumigatus. J Clin Microbiol. 1993 Jun;31(6):1547–1554. doi: 10.1128/jcm.31.6.1547-1554.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hamilton A. J., Holdom M. D., Hay R. J. Specific recognition of purified Cu,Zn superoxide dismutase from Aspergillus fumigatus by immune human sera. J Clin Microbiol. 1995 Feb;33(2):495–496. doi: 10.1128/jcm.33.2.495-496.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hearn V. M., Wilson E. V., Mackenzie D. W. Analysis of Aspergillus fumigatus catalases possessing antigenic activity. J Med Microbiol. 1992 Jan;36(1):61–67. doi: 10.1099/00222615-36-1-61. [DOI] [PubMed] [Google Scholar]
  11. Humphrey T., Sadhale P., Platt T., Proudfoot N. Homologous mRNA 3' end formation in fission and budding yeast. EMBO J. 1991 Nov;10(11):3503–3511. doi: 10.1002/j.1460-2075.1991.tb04914.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Innis M. A., Holland M. J., McCabe P. C., Cole G. E., Wittman V. P., Tal R., Watt K. W., Gelfand D. H., Holland J. P., Meade J. H. Expression, Glycosylation, and Secretion of an Aspergillus Glucoamylase by Saccharomyces cerevisiae. Science. 1985 Apr 5;228(4695):21–26. doi: 10.1126/science.228.4695.21. [DOI] [PubMed] [Google Scholar]
  13. Izard J. W., Kendall D. A. Signal peptides: exquisitely designed transport promoters. Mol Microbiol. 1994 Sep;13(5):765–773. doi: 10.1111/j.1365-2958.1994.tb00469.x. [DOI] [PubMed] [Google Scholar]
  14. Jaton-Ogay K., Paris S., Huerre M., Quadroni M., Falchetto R., Togni G., Latgé J. P., Monod M. Cloning and disruption of the gene encoding an extracellular metalloprotease of Aspergillus fumigatus. Mol Microbiol. 1994 Dec;14(5):917–928. doi: 10.1111/j.1365-2958.1994.tb01327.x. [DOI] [PubMed] [Google Scholar]
  15. Jaton-Ogay K., Suter M., Crameri R., Falchetto R., Fatih A., Monod M. Nucleotide sequence of a genomic and a cDNA clone encoding an extracellular alkaline protease of Aspergillus fumigatus. FEMS Microbiol Lett. 1992 Apr 15;71(2):163–168. doi: 10.1016/0378-1097(92)90506-j. [DOI] [PubMed] [Google Scholar]
  16. Julius D., Brake A., Blair L., Kunisawa R., Thorner J. Isolation of the putative structural gene for the lysine-arginine-cleaving endopeptidase required for processing of yeast prepro-alpha-factor. Cell. 1984 Jul;37(3):1075–1089. doi: 10.1016/0092-8674(84)90442-2. [DOI] [PubMed] [Google Scholar]
  17. Kawasaki L., Wysong D., Diamond R., Aguirre J. Two divergent catalase genes are differentially regulated during Aspergillus nidulans development and oxidative stress. J Bacteriol. 1997 May;179(10):3284–3292. doi: 10.1128/jb.179.10.3284-3292.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kikuchi-Torii K., Hayashi S., Nakamoto H., Nakamura S. Properties of Aspergillus niger catalase. J Biochem. 1982 Nov;92(5):1449–1456. doi: 10.1093/oxfordjournals.jbchem.a134069. [DOI] [PubMed] [Google Scholar]
  19. Kurup V. P., Kumar A. Immunodiagnosis of aspergillosis. Clin Microbiol Rev. 1991 Oct;4(4):439–456. doi: 10.1128/cmr.4.4.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Levitz S. M., Diamond R. D. A rapid colorimetric assay of fungal viability with the tetrazolium salt MTT. J Infect Dis. 1985 Nov;152(5):938–945. doi: 10.1093/infdis/152.5.938. [DOI] [PubMed] [Google Scholar]
  22. Levitz S. M., Diamond R. D. Mechanisms of resistance of Aspergillus fumigatus Conidia to killing by neutrophils in vitro. J Infect Dis. 1985 Jul;152(1):33–42. doi: 10.1093/infdis/152.1.33. [DOI] [PubMed] [Google Scholar]
  23. López-Medrano R., Ovejero M. C., Calera J. A., Puente P., Leal F. An immunodominant 90-kilodalton Aspergillus fumigatus antigen is the subunit of a catalase. Infect Immun. 1995 Dec;63(12):4774–4780. doi: 10.1128/iai.63.12.4774-4780.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. López-Medrano R., Ovejero M. C., Calera J. A., Puente P., Leal F. Immunoblotting patterns in the serodiagnosis of aspergilloma: antibody response to the 90kDa Aspergillus fumigatus antigen. Eur J Clin Microbiol Infect Dis. 1996 Feb;15(2):146–152. doi: 10.1007/BF01591488. [DOI] [PubMed] [Google Scholar]
  25. Miller R. A., Britigan B. E. Role of oxidants in microbial pathophysiology. Clin Microbiol Rev. 1997 Jan;10(1):1–18. doi: 10.1128/cmr.10.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Monod M., Paris S., Sanglard D., Jaton-Ogay K., Bille J., Latgé J. P. Isolation and characterization of a secreted metalloprotease of Aspergillus fumigatus. Infect Immun. 1993 Oct;61(10):4099–4104. doi: 10.1128/iai.61.10.4099-4104.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Monod M., Paris S., Sarfati J., Jaton-Ogay K., Ave P., Latgé J. P. Virulence of alkaline protease-deficient mutants of Aspergillus fumigatus. FEMS Microbiol Lett. 1993 Jan 1;106(1):39–46. doi: 10.1111/j.1574-6968.1993.tb05932.x. [DOI] [PubMed] [Google Scholar]
  28. Navarro R. E., Stringer M. A., Hansberg W., Timberlake W. E., Aguirre J. catA, a new Aspergillus nidulans gene encoding a developmentally regulated catalase. Curr Genet. 1996 Mar;29(4):352–359. [PubMed] [Google Scholar]
  29. Paris S., Monod M., Diaquin M., Lamy B., Arruda L. K., Punt P. J., Latgé J. P. A transformant of Aspergillus fumigatus deficient in the antigenic cytotoxin ASPFI. FEMS Microbiol Lett. 1993 Jul 15;111(1):31–36. doi: 10.1111/j.1574-6968.1993.tb06357.x. [DOI] [PubMed] [Google Scholar]
  30. Perlman D., Halvorson H. O. A putative signal peptidase recognition site and sequence in eukaryotic and prokaryotic signal peptides. J Mol Biol. 1983 Jun 25;167(2):391–409. doi: 10.1016/s0022-2836(83)80341-6. [DOI] [PubMed] [Google Scholar]
  31. Reid T. J., 3rd, Murthy M. R., Sicignano A., Tanaka N., Musick W. D., Rossmann M. G. Structure and heme environment of beef liver catalase at 2.5 A resolution. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4767–4771. doi: 10.1073/pnas.78.8.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rex J. H., Bennett J. E., Gallin J. I., Malech H. L., Melnick D. A. Normal and deficient neutrophils can cooperate to damage Aspergillus fumigatus hyphae. J Infect Dis. 1990 Aug;162(2):523–528. doi: 10.1093/infdis/162.2.523. [DOI] [PubMed] [Google Scholar]
  33. Schroeder W. A., Shelton J. R., Shelton J. B., Robberson B., Apell G., Fang R. S., Bonaventura J. The complete amino acid sequence of bovine liver catalase and the partial sequence of bovine erythrocyte catalase. Arch Biochem Biophys. 1982 Mar;214(1):397–421. doi: 10.1016/0003-9861(82)90044-3. [DOI] [PubMed] [Google Scholar]
  34. Schønheyder H., Andersen P., Petersen J. C. Rapid immunoelectrophoretic assay for detection of serum antibodies to Aspergillus fumigatus catalase in patients with pulmonary aspergillosis. Eur J Clin Microbiol. 1985 Jun;4(3):299–303. doi: 10.1007/BF02013657. [DOI] [PubMed] [Google Scholar]
  35. Tatsumi H., Murakami S., Tsuji R. F., Ishida Y., Murakami K., Masaki A., Kawabe H., Arimura H., Nakano E., Motai H. Cloning and expression in yeast of a cDNA clone encoding Aspergillus oryzae neutral protease II, a unique metalloprotease. Mol Gen Genet. 1991 Aug;228(1-2):97–103. doi: 10.1007/BF00282453. [DOI] [PubMed] [Google Scholar]
  36. Togni G., Sanglard D., Quadroni M., Foundling S. I., Monod M. Acid proteinase secreted by Candida tropicalis: functional analysis of preproregion cleavages in C. tropicalis and Saccharomyces cerevisiae. Microbiology. 1996 Mar;142(Pt 3):493–503. doi: 10.1099/13500872-142-3-493. [DOI] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tran Van Ky P., Biguet J., Vaucelle T. Etude d'une fraction antigénique d'Aspergillus fumigatus support d'une ctivité catalasique. Conséquence sur le diagnostic immunologique de l'aspergillose. Rev Immunol Ther Antimicrob. 1968 Jan-Mar;32(1):37–52. [PubMed] [Google Scholar]
  39. Wayne L. G., Diaz G. A. A double staining method for differentiating between two classes of mycobacterial catalase in polyacrylamide electrophoresis gels. Anal Biochem. 1986 Aug 15;157(1):89–92. doi: 10.1016/0003-2697(86)90200-9. [DOI] [PubMed] [Google Scholar]
  40. Woodbury W., Spencer A. K., Stahman M. A. An improved procedure using ferricyanide for detecting catalase isozymes. Anal Biochem. 1971 Nov;44(1):301–305. doi: 10.1016/0003-2697(71)90375-7. [DOI] [PubMed] [Google Scholar]
  41. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. von Ossowski I., Mulvey M. R., Leco P. A., Borys A., Loewen P. C. Nucleotide sequence of Escherichia coli katE, which encodes catalase HPII. J Bacteriol. 1991 Jan;173(2):514–520. doi: 10.1128/jb.173.2.514-520.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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