Secreted metalloprotease gene family of Microsporum canis

[en] Keratinolytic proteases secreted by dermatophytes are likely to be virulence-related factors. Microsporum canis, the main agent of dermatophytosis in dogs and cats, causes a zoonosis that is frequently reported. Using Aspergillus fumigatus metalloprotease genomic sequence (MEP) as a probe, three genes (MEP1, MEP2, and MEP3) were isolated from an M. canis genomic library. They presented a quite-high percentage of identity with both A. fumigatus MEP and Aspergillus oryzae neutral protease I genes. At the amino acid level, they all contained an HEXXH consensus sequence, confirming that these M. canis genes (MEP genes) encode a zinc-containing metalloprotease gene family. Furthermore, MEP3 was found to be the gene encoding a previously isolated M. canis 43.5-kDa keratinolytic metalloprotease, and was successfully expressed as an active recombinant enzyme in Pichia pastoris. Reverse transcriptase nested PCR performed on total RNA extracted from the hair of M. canis-infected guinea pigs showed that at least MEP2 and MEP3 are produced during the infection process. This is the first report describing the isolation of a gene family encoding potential virulence-related factors in dermatophytes.

Disciplines :

Immunology & infectious disease

Author, co-author :

Brouta, Frédéric

Descamps, Frédéric

Monod, Michel

Vermout, Sandy

Losson, Bertrand ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Mignon, Bernard ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Parasitologie et pathologie des maladies parasitaires

Language :

English

Title :

Secreted metalloprotease gene family of Microsporum canis

Publication date :

October 2002

Journal title :

Infection and Immunity

ISSN :

0019-9567

eISSN :

1098-5522

Publisher :

Amer Soc Microbiology, Washington, United States - Washington

Volume :

Issue :

Pages :

5676-5683

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 17 March 2009

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Bibliography

Apodaca G., McKerrow J.H. (1989) Purification and characterization of a 27,000-Mr extracellular proteinase from Trichophyton rubrum. Infect. Immun. 57:3072-3080.
Apodaca G., McKerrow J.H. (1989) Regulation of Trichophyton rubrum proteolytic activity. Infect. Immun. 57:3081-3090.
Asahi M., Lindquist R., Fukuyama K., Apodaca G., Epstein W.L., McKerrow J.H. (1985) Purification and characterization of major extracellular proteinases from Trichophyton rubrum. Biochem. J. 232:139-144.
Borg-von Zepelin M., Beggah S., Boggian K., Sanglard D., Monod M. (1998) The expression of the secreted aspartyl proteinases Sap4 to Sap6 from Candida albicans in murine macrophages. Mol. Microbiol. 28:543-554.
Brouta F., Descamps F., Fett T., Losson B., Gerday C., Mignon B. (2001) Purification and characterization of a 43.5 kDa keratinolytic metalloprotease from Microsporum canis. Med. Mycol. 39:269-275.
Chambers S.P., Prior S.E., Barstow D.A., Minton N.P. (1988) The pMTL nic- cloning vectors. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene 68:139-149.
Coffey A., Van den Burg B., Veltman R., Abee T. (2000) Characteristics of the biologically active 35-kDa metalloprotease virulence factor from Listeria monocytogenes. J. Appl. Microbiol. 88:132-141.
Collins J.P., Grappel S.F., Blank F. (1973) Role of keratinases in dermatophytosis. II. Fluorescent antibody studies with keratinase II of Trichophyton mentagrophytes. Dermatologica 146:95-100.
De Bernardis F., Arancia S., Morelli L., Hube B., Sanglard D., Schafer W., Cassone A. (1999) Evidence that members of the secretory aspartyl proteinase gene family, in particular SAP2, are virulence factors for Candida vaginitis. J. Infect. Dis. 179:201-208.
Doumas A., Crameri R., Léchenne B., Monod M. (1999) Cloning of the gene encoding neutral protease I of the koji mold Aspergillus oryzae and its expression in Pichia pastoris. J. Food Mycol. 2:271-279.
Duesbery N.S., Webb C.P., Leppla S.H., Gordon V.M., Klimpel K.R., Copeland T.D., Ahn N.G., Oskarsson M.K., Fukasawa K., Paull K.D., Vande Woude G.F. (1998) Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor. Science 280:734-737.
Girardin H., Latgé J.-P. (1994) DNA extraction and quantitation. Molecular biology of pathogenic fungi: a laboratory manual , B. Maresca and G. S. Kobayashi (ed.). Telos Press, New York, N.Y.; 5-9.
Grappel S.F., Blank F. (1972) Role of keratinases in dermatophytosis. I. Immune responses of guinea pigs infected with Trichophyton mentagrophytes and guinea pigs immunized with keratinases. Dermatologica 145:245-255.
Grossberger D. (1987) Minipreps of DNA from bacteriophage lambda. Nucleic Acids Res. 15:6737.
Jaton-Ogay K., Paris S., Huerre M., Quadroni M., Falchetto R., Togni G., Latgé J.P., Monod M. (1994) Cloning and disruption of the gene encoding an extracellular metalloprotease of Aspergillus fumigatus. Mol. Microbiol. 14:917-928.
Jaton-Ogay K., Suter M., Crameri R., Falchetto R., Fatih A., Monod M. (1992) Nucleotide sequence of a genomic and cDNA clone encoding an extracellular alkaline protease of Aspergillus fumigatus. FEMS Microbiol. Lett. 92:163-168.
Katz M.E., Rice R.N., Cheetham B.F. (1994) Isolation and characterization of an Aspergillus nidulans gene encoding an alkaline protease. Gene 150:287-292.
Laemmli U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685.
Lambkin I., Hamilton A.J., Hay R.J. (1996) Purification and characterisation of a novel 34,000-Mr cell-associated proteinase from the dermatophyte Trichophyton rubrum. FEMS Immunol. Med. Microbiol. 13:131-140.
Lateur N. (2000) Tinea capitis chez l'enfant: Diagnostic, étiologie et traitement. Percentile 5:100-103.
Lee K.H., Park H.W., Lee J.B. (1991) Localization of keratinolytic proteinase in skin tissue sections of guinea pigs with Microsporum canis infection by immunoperoxidase technique in electron microscopy. Ann. Dermatol. 3:1-4.
Lee K.H., Park K.K., Park S.H., Lee J.B. (1987) Isolation, purification and characterization of keratinolytic proteinase from Microsporum canis. Yonsei Med. J. 28:131-138.
Lunder M., Lunder M. (1992) Is Microsporum canis infection about to become a serious dermatological problem?. Dermatology 184:87-89.
Markaryan A., Morozova I., Yu H., Kolattukudy P.E. (1994) Purification and characterization of an elastinolytic metalloprotease from Aspergillus fumigatus and immunoelectron microscopic evidence of secretion of this enzyme by the fungus invading the murine lung. Infect. Immun. 62:2149-2157.
Meevootisom V., Niederpruem D.J. (1979) Control of exocellular proteases in dermatophytes and especially Trichophyton rubrum. Sabouraudia 17:91-106.
Mignon B., Coignoul F., Leclipteux T., Focant C., Losson B. (1999) Histopathological pattern and humoral immune response to a crude exoantigen and purified keratinase of Microsporum canis in symptomatic and asymptomatic infected cats. Med. Mycol. 37:1-9.
Mignon B., Leclipteux T., Focant C., Nikkels A., Pierard G., Losson B. (1999) Humoral and cellular immune response to a crude exo-antigen and purified keratinase of Microsporum canis in experimentally infected guinea pigs. Med. Mycol. 37:123-129.
Mignon B., Nikkels A., Pierard G., Losson B. (1998) The in vitro and in vivo production of a 31.5 kDa keratinolytic subtilase from Microsporum canis and the clinical status in naturally infected cats. Dermatology 196:438-441.
Mignon B., Swinnen M., Bouchara J.P., Hofinger M., Nikkels A., Pierard G., Gerday C., Losson B. (1998) Purification and characterization of a 31.5 kDa keratinolytic subtilisin-like serine protease from Microsporum canis and evidence of its secretion in naturally infected cats. Med. Mycol. 36:395-404.
Miyoshi S., Shinoda S. (2000) Microbial metalloproteases and pathogenesis. Microbes Infect. 2:91-98.
Monod M., Paris S., Sanglard D., Jaton-Ogay K., Bille J., Latgé J.P. (1993) Isolation and characterization of a secreted metalloprotease of Aspergillus fumigatus. Infect. Immun. 61:4099-4104.
Monod M., Togni G., Hube B., Sanglard D. (1994) Multiplicity of genes encoding secreted aspartic proteinases in Candida species. Mol. Microbiol. 13:357-368.
Naglik J.R., Newport G., White T.C., Fernandes-Naglik L.L., Greenspan J.S., Greenspan D., Sweet S.P., Challacombe S.J., Agabian N. (1999) In vivo analysis of secreted aspartyl proteinase expression in human oral candidiasis. Infect. Immun. 68:2482-2490.
Okeke C.N., Tsuboi R., Kawai M., Hiruma M., Ogawa H. (2001) Isolation of an intron-containing partial sequence of the gene encoding dermatophyte actin (ACT) and detection of a fragment of the transcript by reverse transcription-nested PCR as a means of assessing the viability of dermatophytes in skin scales. J. Clin. Microbiol. 39:101-106.
Ramesh M.V., Sirakova T., Kolattukudy P.E. (1994) Isolation, characterization, and cloning of cDNA and the gene for an elastinolytic serine proteinase from Aspergillus flavus. Infect. Immun. 62:79-85.
Rawlings N.D., Barrett A.J. (1995) Evolutionary families of metallopeptidases. Methods Enzymol. 248:183-228.
Rawlings N.D., Barrett A.J. (2000) MEROPS: The peptidase database. Nucleic Acids Res. 28:323-325.
Romano C. (1999) Tinea capitis in Siena, Italy. An 18-year survey. Mycoses 42:559-562.
Sambrook J., Fritsch E.F., Maniatis T. Molecular cloning: a laboratory manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.; 1989.
Schaller M., Korting H.C., Schafer W., Bastert J., Cen W., Hube B. (1999) Secreted aspartic proteinase (Sap) activity contributes to tissue damage in a model of human oral candidosis. Mol. Microbiol. 34:169-180.
Scott D.W., Miller W.H., Griffin C.E. (1995) Fungal skin diseases. Small animal dermatology, 5th ed. , G. H. Muller (ed.). W. B. Saunders, Philadelphia, Pa.; 329-391.
Takiuchi I., Higuchi D., Sei Y., Koga K. (1982) Isolation of an extra-cellular proteinase (keratinase) from Microsporum canis. Sabouraudia 20:281-288.
Takiuchi I., Sei Y., Tagaki H., Negi M. (1984) Partial characterization of the extracellular keratinase from Microsporum canis. Sabouraudia 22:219-224.
Tatsumi H., Ohsawa M., Tsuji R.F., Murakami S., Nakano E., Motai H., Masaki A., Ishida Y., Murakami K., Kawabe H., Arimura H. (1988) Cloning and sequence of the alkaline protease cDNA from Aspergillus oryzae. Agric. Biol. Chem. 52:1887-1888.
Towbin H., Staehelin T., Gordon J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. USA 76:4350-4354.
Tsuboi R., Ko I.J., Takamori K., Ogawa H. (1989) Isolation of a keratinolytic proteinase from Trichophyton mentagrophytes with enzymatic activity at acidic pH. Infect. Immun. 57:3479-3483.
Van Cutsem J. (1989) Animal models for dermatomycotic infections. Current topics in medical mycology , M. R. McGinnis and M. Borgers (ed.). Springer-Verlag, New York, N.Y.; 3:1-35.
Weitzman I., Summerbell R.C. (1995) The dermatophytes. Clin. Microbiol. Rev. 8:240-259.
Woodfolk J.A., Wheatley L.M., Piyasena R.V., Benjamin D.C., Platts-Mills T.A. (1998) Trichophyton antigens associated with IgE antibodies and delayed type hypersensitivity. Sequence homology to two families of serine proteinases. J. Biol. Chem. 273:29489-29496.
Proceedings of the ISHAM consultation on prevention and control of dermatophytosis with special reference to immunity and immunization , National Veterinary Institute, Oslo, Norway; 1987, 1-40.
Yu R.J., Harmon S.R., Blank F. (1968) Isolation and purification of an extracellular keratinase of Trichophyton mentagrophytes. J. Bacteriol. 96:1435-1436.
Yu R.J., Harmon S.R., Grappel S.F., Blank F. (1971) Two cell-bound keratinases of Trichophyton mentagrophytes. J. Investig. Dermatol. 56:27-32.
Zaugg C., Borg-Von Zepelin M., Reichard U., Sanglard D., Monod M. (2001) Secreted aspartic proteinase family of Candida tropicalis. Infect. Immun. 69:405-412.