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Amino Acid Alterations in the $\beta$- Tubulin of Metarhizium anisopliae That Confer Benomyl Resistance  

Kim Soon Kee (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Shim Hee Jin (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Roh Jong Yul (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Li Ming Shun (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Choi Jae Young (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Jin Byung Rae (College of Natural Resources and Life Science, Dong-A University)
Boo Kyung Saeng (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Je Yeon Ho (School of Agricultural Biotechnology, College of Agriculture & Life Sciences, Seoul National University)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.11, no.1, 2005 , pp. 37-42 More about this Journal
Abstract
We cloned the $\beta$-tubulin genes from the wild type strain and two benomyl-resistant mutants of Metahizium anisopliae and determined their nucleotide sequences. A $\beta$-tubulin encoding 448-residue protein from wild type M. anisopliae shows strong homology to other $\beta$-tubulins. The coding region is interrupted by four introns. Comparisons of intron position between the M. anisopliae gene and other fungal $\beta$-tubulin genes show considerable positional conservation. The mutations responsible for benomyl resistance were determined in two spontaneous mutants, 8-18 and 8­19. One mutant 8-18 substituted glutamate for aspar­agine at position 33 and lysine for glutamine at position 134. The other mutant 8-19 showed alterations at three positions of $\beta$-tubulin arginine for tryptophan at position 21, lysine for asparagine at position 33, and phenylalanine for leucine at position 240. These data suggest that regions of $\beta$-tubulin containing amino acids 21, 33,134, and 240 interact to form the binding site of benomyl.
Keywords
Benomyl; beta-tubulin gene; Entomopathogenic fungus; Metahizium anisopliae;
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  • Reference
1 Deacon, J. W. (1997) Modem mycology. pp. 278-281, Blackwell Science, Fitchburg
2 Delp, C. J. (1995). Benzimidazole and related fungicides; in Modern Selective Fungicides. Lyr, H. (ed.), pp. 291-303, Gustv Fisher Verlag, Jena
3 Jung, M. K., I. B. Isaac and B. R. Oakley (1992) Amino acid alterations in the benA gene of Aspergillus nidulans that confer benomyl resistance. Cell Motil. Cytoskel. 22, 170-174   DOI   ScienceOn
4 Moore, D. and C. Prior (1993) The potential of mycoinsecticides. Biocon. News Inform. 14, 3IN-40N
5 Davidse, L. C. (1986) Benzimidazole fungicides: mechanism of action and biological impact. Annu. Rev. Phytopathol. 24, 43-65   DOI   ScienceOn
6 Boucias, D. G and J. C. Pend land (1998) Principles of insect pathology. Kluwer Academic Publishers, Massachusetts
7 Langford, C. J., F. J. Klinz, C. Donath and D. Gallwitz (1984) Point mutations identify the conserved, intron-contained TACTAAC box as an essential splicing signal sequence in yeast. Cell 36, 645-653   DOI   ScienceOn
8 Seip, E. R., C. P. Woloshuk, G. A. Payne and S. E. Curtis (1990) Isolation and sequence analysis of a $\beta$-tubulin gene from Aspergillus flavus and its use as a selectable marker. Appl. Envron. Microbiol. 56,3686-3692
9 Davidse, L. C. and H. Ishii (1995) Biochemical and molecular aspects of the mechanisms of action of benzimidazoles, Nphenylcarbamates and N-phenylformadoximes and the mechanisms of resistance to these compounds in fungi; in Modern Selective Fungicides. Lyr, H. (ed.), pp. 305-322, Gustv Fisher Verlag, Jena
10 Nogales, E., S. G. Wolf and K. H. Downing (1998) Structure of the ${\alpha}{\beta}$ tubulin dimer by electron crystallography. Nature 391, 199-203   DOI   PUBMED   ScienceOn
11 May, G. S., L. S. Tsang, H. Smith, S. Fidel and R.N. Morris (1987) Aspergillus nidulans $\beta$-tubulin genes are unusually divergent. Gene 55, 231-243   DOI   PUBMED   ScienceOn
12 Samish, M. and J. Rehacek (1999) Pathogens and predators of ticks and their potential in biological control. Annu. Rev. Entomol. 44, 159-182   DOI   ScienceOn
13 Davidse, L. C. and W. Flach (1977) Differential binding of methyl benzimidazol-2-yl carbamate to fungal tubulin as a mechanism of resistance to this antimitotic agent in mutant strains of Aspergillus nidulans. J. Cell Biol. 72, 174-193   DOI   ScienceOn
14 Goettel, M. S., R. J. St. Leger, S. Bhairi, M. K. Jung, B. R. Oakley, D. W. Roberts and R. C. Staples (1990) Pathogenicity and growth of Metarhizium anisopliae stably transformed to benomyl resistance. Curr. Genet. 17, 129-132   DOI
15 Smith, H. A., H. S. Allaudeen, M. H. Whitman, Y. Koltin and A. Gorman (1988) Isolation and characterization of a $\beta$-tubulin gene from Candida albicans. Gene 63,53-63   DOI   PUBMED   ScienceOn
16 Jung, M. K. and B. R. Oakley (1990) Identification of an amino acid substitution in the benA, $\beta$-tubulin gene of Aspergillus nidulans that confers thiabendazole resistance and benomyl supersensitivity. Cell Motil. Cytoskel. 17, 87-94   DOI   ScienceOn
17 Orbach, M. J., E. B. Porro and C. Y. Yanofsky (1986) Cloning and characterization of the gene for $\beta$-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker. Mol. Cell Biol. 6, 2452-2461
18 Sheir-Neiss, G., M. H. Lai and N. R. Morris (1978) Identification of a gene for $\beta$-tubulin gene in Aspergillus nidulans. Cell 15, 639-647   DOI   PUBMED   ScienceOn
19 Butters, J. A., K. U. Devi, C. M. Mohan and V. Sridevi (2003) Screening for tolerance to bavistan, a benzimidazole fungicide containing methyl benimidazol-2-yl carbamate (MBC), in Beauveria bassiana: Sequence analysis of the $\beta$-tubulin gene to identify mutations conferring tolerance. Mycol. Res. 107, 260-266   DOI   ScienceOn
20 Sambrook, L, E. F. Fritsch and T. Maniatis (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New York
21 Thomas, J. H., N. Neff and D. Dotstein (1985) Isolation and characterization of mutations in the $\beta$-tubulin gene of Saccharomyces cereviciae. Genetics 111, 715-734
22 Mckay, G. J., D. Egan, E. Morris and A. E. Brown (1998) Identification of benzimidazole resistance in Cladobotryum dendroides using a PCR-based method. Mycol. Res. 102, 671-676   DOI   ScienceOn
23 Park, S. Y., O. J. Jung, Y. R. Chung and C. W. Lee (1997) Isolation and characterization of a benornyl-resistant form of -$\beta$tubulin-encoding gene from the phytopathogenic fungus Botryotina fuckeliana. Mol. Cells. 7, 104-109