PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of ijclbiowww.springer.comThis journalToc AlertsSubmit OnlineOpen Choice
 
Indian J Clin Biochem. Jan 2005; 20(1): 208–212.
PMCID: PMC3454157
Identification and characterization of polyubiquitin gene from cDNA library of aspergillus fumigatus
Jata Shankar,corresponding author1 Taruna Madan,corresponding author1 Seemi Farhat Basir,2 and P. Usha Sarma3
1Molecular Biochemistry and Diagnostics Division, Institute of Genomics and Integrative Biology, Council for Scientific and Industrial Research, Mall Road, 110007 Delhi,
2Department of Biosciences, Jamia Millia Islamia, Jamia Nagar, 110025 New Delhi,
3Department of Plant Pathology, Indian Agricultural Research Institute, 110012 Pusa,
Taruna Madan, taruna_m/at/hotmail.com.
corresponding authorCorresponding author.
Abstract
Aspergillus fumigatus (Afu) causes allergic and invasive forms of diseases in humans. In order to identify genes relevant for pathogenesis, a total of 235 cDNA clones were randomly selected and sequenced from cDNA library of Afu. One of the partially sequenced cDNA clones was homologous to polyubiquitin. Sequencing of the complete cDNA clone showed an open reading frame of 912 bases. Comparison with genomic sequence of Afu using BlastN program, revealed that polyubiquitin gene comprises of 992 bases and contains one intron of 80 bases. The recombinant expression of fusion protein showed an approximately molecular weight of 43-kDa on SDS-PAGE. The translation product of the cDNA sequence showed four tandem repeats of 76 amino acid residues in a single polyubiquitin protein and showed 100% identity with polyubiquitin protein sequences of S. cerevisiae, N. crassa, C. albicans, S. pombe, and M. grisae. Polyubiquitin gene is known to play important role in a variety of cellular processes and recently have been implicated in fungal pathogenesis. Identification of polyubiquitin gene of Afu has opened up scope to study its role in understanding Aspergillus biology and pathogenesis.
Full Text
The Full Text of this article is available as a PDF (573K).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
1. Pitt J.I., Samson R.A., Frisvad J.C. Integration of modern taxonomic methods for Penicillium and Aspergillus classification. Amsterdam: Harwood Publishers; 2000. pp. 9–49.
2. Yu J., Bhatnagar D., Ehrlich K.C. Aflatoxin biosynthesis. Rev. Iberoam. Micol. 2002;19:191–200. [PubMed]
3. Latge J.P. Aspergillus fumigatus and aspergillosis. Clin. Microbial. Rev. 1999;12:310–350. [PMC free article]
4. Heather R.K., McCafferty, Talbot N.J. Identification of three ubiquitin genes of the rice blast fungus Magnaporthe grisea, one of which is highly expressed during initial stages of plant colonization. Curr. Genet. 1998;33:352–361. doi: 10.1007/s002940050347. [PubMed] [Cross Ref]
5. Kurokawa C.S., Sugizaki M.F., Peracoli M.T. Virulent factors in fungi of systemic mycoses. Rev. Inst. Med. Trop. Sao. Paula. 1998;40:125–135.
6. Fillingham J.S., Chilcoat N.D., Turkewitz A.P., Orias E., Reith M., Pearlman R.E. Analysis of expressed sequence tags (ESTs) in the ciliated protozoan Tetrahymena thermophila. J. Eukaryot. Microbiol. 2002;49:99–107. doi: 10.1111/j.1550-7408.2002.tb00350.x. [PubMed] [Cross Ref]
7. Gouwu W., Xinbing Y.U., Zhongdaola W.U. Generation and analysis of 113 adult stage Schistosoma japonicum (chinese strain) expressed sequence tags. Chin. Med. J. 2002;115:1517–1520. [PubMed]
8. Neumann M.J., Dobinson K.F. Sequence tag analysis of gene expression during pathogenic growth and microsclerotia development in the vascular wilt pathogen Verticillium dahliae. Fungal Genet. Bio. 2002;38:54–62. doi: 10.1016/S1087-1845(02)00507-8. [PubMed] [Cross Ref]
9. Verdun R.E., Dipaloo N., Urmenyi T.P., Rondinelli E., Frasch A.C., Sanchez D.O. Gene discovery through expressed sequence tag sequencing in Trypanosoma cruzi. Infec. Immun. 1998;66:5393–5398. [PMC free article] [PubMed]
10. Shankar J., Nigam S., Saxena S., Madan T., Sarma P.U. Identification and Assignment of Function to the Genes of Aspergillus fumigatus Expressed at 37°C. J. Eukaryot. Microbiol. 2004;51:428–432. doi: 10.1111/j.1550-7408.2004.tb00390.x. [PubMed] [Cross Ref]
11. Maneger I.D., Hehl A., Parmeley S., Sibley L.D., Marra M., Hiller L., Waterson R., Boothroyd J.C. Expressed sequence tag analysis of the bradyzoite stage of Toxoplasma gondii: Identification of developmentally regulated genes. Infec. Immun. 1998;66:1632–1637. [PMC free article] [PubMed]
12. Goldman G.H., Marques E.D.R., Ribeiro D.C.D., Bernardes L.A.D.S., Quiapin A.C., Vitorelli P.M., et al. Expressed sequence tag analysis of the human pathogen Paraceccidioides brasiliensis yeast phase: Identification of putative homologues of Candida albicans virulence and pathogenicity genes. Eukaryot. Cell. 2003;2:34–48. doi: 10.1128/EC.2.1.34-48.2003. [PMC free article] [PubMed] [Cross Ref]
13. Saxena S., Madan T., Muralidhar K., Sarma P.U. cDNA cloning, expression and characterization of an allergenic L3 ribosomal protein of Aspergillus fumigatus. Clin. Exp. Immunol. 2003;134:86–91. doi: 10.1046/j.1365-2249.2003.02257.x. [PubMed] [Cross Ref]
14. Nigam S., Sarma P.V.G.K., Ghosh P.C., Sarma P.U. Characterization of Aspergillus fumigatus protein disulfide isomerase family gene. Gene. 2001;281:143–50. doi: 10.1016/S0378-1119(01)00794-6. [PubMed] [Cross Ref]
15. Sambrook J., Fritsch E.F., Maniatis T. “Molecular cloning a laboratory manual”. Second Ed. New York: Cold Spring harbour; 1989.
16. Jentsch S. The ubiquitin-conjugation systeM. Ann. Rev. Genet. 1992;26:179–207. doi: 10.1146/annurev.ge.26.120192.001143. [PubMed] [Cross Ref]
17. Hershko A., Ciechanover A. The ubiquitin system. Annu. Rev. Biochem. 1998;67:425–479. doi: 10.1146/annurev.biochem.67.1.425. [PubMed] [Cross Ref]
18. Ciechanover A. The ubiquitin-proteasome pathway: on protein death and cell life. EMBO J. 1998;24:7161–7160.
19. Finley D., Bartel B., Varshavsky A. The tails of ubiquitin precursors are ribosomal proteins whose fusion to ubiquitin facilitates ribosome biogenesis. Nature. 1989;338:394–401. doi: 10.1038/338394a0. [PubMed] [Cross Ref]
20. Sepulveda P., Lopez-Ribot J.L., Gozalbo D., Cervera A., Martinez J.P., Chaffin W.L. Ubiquitin-like epitopes associated with Candida albicans cell surface receptors. Infect. Immun. 1996;64:4406–4408. [PMC free article] [PubMed]
21. Bouchara J.P., Sanchez M., Chevailler A., Marot-Leblond A., Lissitzky J.C., Tronchin G., Chabasse D. Sialic acid-dependent recognition of laminin and fibrinogen by Aspergillus fumigatus conidia. Infect Immun. 1997;65:2717–2724. [PMC free article] [PubMed]
22. Tronchin G., Esnault K., Renier G., Filmon R., Chabasse D., Bouchara J.P. Expression and identification of a laminin-binding protein in Aspergillus fumigatus conidia. Infect. Immun. 1997;65:9–15. [PMC free article] [PubMed]
Articles from Indian Journal of Clinical Biochemistry are provided here courtesy of
Springer