Aspergillus fumigatus alb1 encodes naphthopyrone synthase when expressed in Aspergillus oryzae
Akira Watanabe
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Search for more papers by this authorIsao Fujii
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Search for more papers by this authorHuei-Fung Tsai
Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1882, USA
Search for more papers by this authorYun C. Chang
Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1882, USA
Search for more papers by this authorK.J. Kwon-Chung
Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1882, USA
Search for more papers by this authorCorresponding Author
Yutaka Ebizuka
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
*Corresponding author. Tel.: +81 (3) 5841-4740; Fax: +81 (3) 5841-4744, E-mail address: [email protected]Search for more papers by this authorAkira Watanabe
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Search for more papers by this authorIsao Fujii
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Search for more papers by this authorHuei-Fung Tsai
Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1882, USA
Search for more papers by this authorYun C. Chang
Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1882, USA
Search for more papers by this authorK.J. Kwon-Chung
Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1882, USA
Search for more papers by this authorCorresponding Author
Yutaka Ebizuka
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
*Corresponding author. Tel.: +81 (3) 5841-4740; Fax: +81 (3) 5841-4744, E-mail address: [email protected]Search for more papers by this authorAbstract
Conidial pigment biosynthesis is an important virulence factor in Aspergillus fumigatus, a human fungal pathogen. Involvement of DHN–melanin pathway in the biosynthesis of A. fumigatus conidial pigment implies that the Alb1p polyketide synthase (PKS) is a 1,3,6,8-tetrahydroxynaphthalene (T4HN) synthase. The Alb1p, however, shows higher sequence similarity to a naphthopyrone synthase than to a T4HN synthase. To clarify the function of Alb1p, the alb1 gene was overexpressed in a heterologous host Aspergillus oryzae. The Alb1p PKS product in this heterologous expression system was identified as heptaketide naphthopyrone instead of pentaketide T4HN. The data suggest that Alb1p is a naphthopyrone synthase.
References
- [1] Kwon-Chung, K.J. and Bennett, J.E. (1992) Aspergillosis. In: Medical Mycology, pp. 201–247. Williams and Wilkins, Baltimore, MD.
- [2] Cohen, M.S., Isturiz, R.E., Malech, H.L., Root, R.K., Wilfert, C.M., Gutman, L., Buckley, R.H. (1981) Fungal infection in chronic granulomatous disease: the importance of the phagocyte in defence against fungi. Am. J. Med. 71, 59–66.
- [3] Denning, D.W. (1996) Therapeutic outcome in invasive aspergillosis. Clin. Infect. Dis. 23, 608–615.
- [4] Gerson, S.T., Talbot, G.H., Hurwitz, S., Strom, B.L., Lusk, E.J., Cassileth, P.A. (1984) Prolonged granulocytopenia: the major risk factor for invasive pulmonary aspergillosis in patients with acute leukemia. Ann. Intern. Med. 100, 345–351.
- [5] Gustafson, T.L., Schaffner, W., Lavely, G.B., Stratton, C.W., Johnson, H.K., Hutcheson, J.R.H. (1983) Invasive aspergillosis in renal transplant recipients: correlation with corticosteroid therapy. J. Infect. Dis. 148, 230–238.
- [6] Wheeler, M.H. and Bell, A.A. (1988) Melanins and their importance in pathogenic fungi. In: Current Topics in Medical Mycology (McGinnis, M.R., Ed.), Vol. 2, pp. 338–387. Springer, New York.
- [7] Bell, A.A., Wheeler, M.H. (1986) Biosynthesis and function of fungal melanins. Annu. Rev. Phytopathol. 24, 411–451.
- [8] Chumley, F.G., Valent, B. (1990) Genetic analysis of melanin-deficient, nopathogenic mutants of Magnaporthe grisea. Mol. Plant-Microbe Interact. 3, 135–143.
- [9] Tayler, B.E., Wheeler, M.H., Szaniszlo, P.J. (1987) Evidence for pentaketide melanin biosynthesis in dematiaceous human pathogenic fungi. Mycologia 79, 320.
- [10] Wheeler, M.H. (1983) Comparison of fungal melanin biosynthesis in ascomycetous, imperfect and basidiomycetes fungi. Trans. Br. Mycol. Soc. 81, 29–36.
- [11] O'Hara, E.B. and Timberlake, W.E. (1989) Molecular characterization of the Aspergillus nidulans yA locus. Genetics 121, 249–254 (Erratum: (1990) 124, 791).
- [12] Aramayo, R., Timberlake, W.E. (1990) Sequence and molecular structure of the Aspergillus nidulans yA (laccase I) gene. Nucleic Acids Res. 18, 3415.
- [13] Mayorga, M.E., Timberlake, W.E. (1990) Isolation and molecular characterization of the Aspergillus nidulans wA gene. Genetics 126, 73–79.
- [14] Mayorga, M.E., Timberlake, W.E. (1992) The developmentally regulated Aspergillus nidulans wA gene encodes a polypeptide homologous to polyketide and fatty acid synthases. Mol. Gen. Genet. 235, 205–212.
- [15] Watanabe, A., Ono, Y., Fujii, I., Sankawa, U., Mayorga, M.E., Timberlake, W.E., Ebizuka, Y. (1998) Product identification of polyketide synthase coded by Aspergillus nidulans wA gene. Tetrahedron Lett. 39, 7733–7736.
- [16] Watanabe, A., Fujii, I., Sankawa, U., Mayorga, M.E., Timberlake, W.E., Ebizuka, Y. (1999) Re-identification of Aspergillus nidulans wA gene to code for a polyketide synthase of naphthopyrone. Tetrahedron Lett. 40, 91–94.
- [17] Tokousbalides, M.C., Sisler, H.D. (1979) Site of inhibition by tricyclazole in the melanin biosynthetic pathway of Verticillium dahliae. Pestic. Biochem. Physiol. 11, 64–73.
- [18] Wheeler, M.H., Klich, M.A. (1995) The effects of tricyclazole, pyroquilon, phthalide, and related fungicides on the production of conidial wall pigments by Penicillium and Aspergillus species. Pestic. Biochem. Physiol. 52, 125–136.
- [19] Tsai, H.-F., Washburn, R.G., Chang, Y.C., Kwon-Chung, K.J. (1997) Aspergillus fumigatus arp1 modulates conidial pigmentation and complement deposition. Mol. Microbiol. 26, 175–183.
- [20] Tsai, H.-F., Wheeler, M.H., Chang, Y.C., Kwon-Chung, K.J. (1999) A developmentally regulated gene cluster involved in conidial pigment biosynthesis in Aspergillus fumigatus. J. Bacteriol. 181, 6469–6477.
- [21] Tsai, H.-F., Chang, Y.C., Washburn, R.G., Wheeler, M.H., Kwon-Chung, K.J. (1998) The developmentally regulated alb1 gene of Aspergillus fumigatus: its role in modulation of conidial morphology and virulence. J. Bacteriol. 180, 3031–3038.
- [22] Fujii, I., Mori, Y., Watanabe, A., Kubo, Y., Tsuji, G., Ebizuka, Y. (1999) Heterologous expression and product identification of Colletotrichum lagenarium polyketide synthase encoded by the PKS1 gene involved in melanin biosynthesis. Biosci. Biotechnol. Biochem. 63, 1445–1452.
- [23] Fujii, I., Ono, Y., Tada, H., Gomi, K., Ebizuka, Y., Sankawa, U. (1996) Cloning of the polyketide synthase gene atX from Aspergillus terreus and its identification as the 6-methylsalicylic acid synthase gene by heterologous expression. Mol. Gen. Genet. 253, 1–10.
- [24] Fujii, T., Yamaoka, H., Gomi, K., Kitamoto, K., Kumagai, C. (1995) Cloning and nucleotide sequence of the ribonuclease T1 Gene (rntA) from Aspergillus oryzae and its expression in Saccharomyces cerevisiae and Aspergillus oryzae. Biosci. Biotechnol. Biochem. 59, 1869–1874.
- [25] Gomi, K., Iimura, Y., Hara, S. (1987) Integrative transformation of Aspergillus oryzae with a plasmid containing the Aspergillus nidulans argB gene. Agric. Biol. Chem. 51, 2549–2555.
- [26] Fujii, I., Watanabe, A., Mori, Y., Ebizuka, Y. (1998) Structures and functional analyses of fungal polyketide synthase genes. Actinomycetology 12, 1–14.
- [27] Fujii, I. (1999) Polyketide biosynthesis in filamentous fungi. In: Comprehensive Natural Products Chemistry (Sankawa, U., Ed.), Vol. 1, pp. 409–441. Elsevier, Oxford.
- [28] Kennedy, J., Auclair, K., Kendrew, S.G., Park, C., Vederas, J.C., Hutchinson, C.R. (1999) Modulation of polyketide synthase activity by accessory proteins during lovastatin biosynthesis. Science 284, 1368–1372.
