Effect of phosphatidylcholine on the level expression of plc genes of Aspergillus fumigatus by real time PCR method and investigation of these genes using bioinformatics analysis
,
Abstract
Background and Objectives: Phosphlipases are a group of enzymes that breakdown phosphatidylcholine (phospholipids) molecules producing second products. These produced products have a divers role in the cell like signal transduction and digestion in humans. In this research the effect of phosphatidylcholine on the expression of plc genes of A. fumigatus was studied. The plc genes of this fungus were also interrogated using bioinformatics studies.
Materials and Methods: Real-time PCR was performed to study the expression of plc genes and these genes were interrogated using bioinformatics studies.
Results: There was more significant expression for all three plc genes when A. fumigatus was grown on the presence of phosphatidylcholine in the medium. The sequence of plc genes of A. fumigatus was also interrogated using bioinformatics analysis and their relationship with the other microorganisms was investigated.
Conclusion: Real-time PCR revealed that afplc1, afplc2 and afplc3 were up-regulated in the presence of phosphatidylcho- line. In this study we suggest either the plc’s of A. fumigatus were present in an ancestral genome and have become lost in some lineages, or that they have been acquired from other organisms by horizontal gene transfer. We also found that plc’s of this fungus appeared to be more closely related to the plant plc’s than the bacterial plc’s.
Rippon, JW. Medical Mycology The Pathogenic Fungi and The Pathogenic Actinomycetes. Philadelphia: B. Saunders Company; 1988.
Franquet T, Gimenez A, Hidalgo A. Imaging of opportunistic fungal infections in immunocompromised patient. Euro J Radio 2004; 51: 130-138.
Torres HA, Rivero GA, Lewis RE, Hachem R, Raad II, Kontoyiannis DP. Aspergillosis caused by non- fumigatus Aspergillus Species: risk factors and in Vitro susceptibility compared with Aspergillus fumigatus. Diag Micro Infect Dis 2003; 46: 25-28.
Li X, Gao M, Han M, Tao S, Zheng D, Cheng Y, et al. Disruption of the Phospholipase D Gene Attenuates the Virulence of Aspergillus fumigatus. Infect Immun 2012; 80: 429-440.
Haines J. Aspergillus in compost: straw man or fatal flaw. Biocylce 1995; 6: 32-35.
Latge J P. Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev 1999; 12: 310-350.
Mellor H, Parker P. The extended protein kinase C superfamily. Biochem J 1998; 332: 281-292.
Newton, AC. Protein kinase C: structural and spatial regulation by phosphorylation, cofactors, and macromolecular interactions. Chem Rev 2001; 101:2353-2364.
Patterson RL, Boehning D, Snyder SH. Inositol 1,4,5- trisphosphate receptors as signal integrators. Annu Rev Biochem 2004; 73: 437-465.
Pattni K, Banting, G. Ins (1, 4, 5) P3 metabolism and the family of IP3-3Kinases. Cell Signal 2004; 16: 643-654.
Schmitz HP, Heinisch JJ. Evolution, biochemistry and genetics of protein kinase C in fungi. Curr Genet 2003;43: 245-254.
Sorrentino V, Rizzuto R. Molecular genetics of Ca2+stores and intracellular Ca2+ signalling. Tren Pharm Sci 2002; 22: 459-464.
Berka RM, Gray GL, Vasil ML. Studies of phospholipase C (heat-labile hemolysin) in Pseudomonas aeruginosa. Infect Immun 1981; 34: 1071-1074.
Gilmore MS, Cruz-Rodz AL, Leimeister-Wachter M, Kreft J, Goebel W. A Bacillus cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and sphingomyelinase genes: nucleotide sequence and genetic linkage. J Bacteriol 1989; 171: 744-753.
Logan AJ, Williamson ED, Titball RW, Percival DA, Shuttleworth AD, Conlan JW, et al. Epitope mapping of the alpha-toxin of Clostridium perfringens. Infect Immun 1991; 59: 4338-4342.
Raynaud C, Guilhot C, Rauzier J, Bordat Y, Pelicic V, Manganelli R, et al. Phospholipases C are involved in the virulence of Mycobacterium tuberculosis. Mol Microbiol 2002; 45: 203-217.
Titball RW, Fearn AM, Williamson ED. Biochemical and immunological properties of the C-terminal domain of the alpha-toxin of Clostridium perfringens. FEMS Microbiol Lett 1993; 110: 45-50.
Vazquez-Boland JA, Kocks C, Dramsi S, Ohayon H, Geoffroy C, Mengaud J, et al. Nucleotide sequence of the lecithinase operon of Listeria monocytogenes and possible role of lecithinase in cell-to-cell spread. Infect Immun 1992; 60: 219-230.
Shen DK, Noodeh AD, Kazemi A, Grillot R, Robson G, Brugère JF. Characterisation and expression of phospholipases B from the opportunistic fungus Aspergillus fumigatus. FEMS Microbiol Lett 2004; Oct 1; 239: 87-93.
Kazemi AH, Robson GD, Identifying phospholipase B molecule as an virulence factor in microorganisms (In Persian Language). Journal of Hamedan Medical Sciences University 2003; 10 : 63-56.
Kazemi AH, Cloning and sequencing a partial of phospholipase B1 gene of A. fumigates (In Persian Language). Journal of Tabriz Medical Sciences University 2004; 38: 47-54.
Kazemi AH, Robson GD, Denning D. Cloning and sequencing a partial of phospholipase B2 gene of A. fumigates (In Persian Language). Komesh 2004; 6: 38-94.
Kazemi AH, Robson GD, Denning D, Zarei- Mahmoudabadi A, Jaffari-NAdoushan AA. Identifying of phospholipase B3 molecule as important factor in the pathogenicity of A. fumigatus (In Persian Language). Journal of Mashhad Medical Sciences University 2010;53: 199-205.
Kazemi AH, Robson GD, Denning D. Identifying of phospholipase D molecule as important factor in the pathogenicity of microorganisms (In Persian Language). Journal of Tabriz Medical Sciences University 2007;15: 35-45.
Birch M, Robson G, Law D, Denning, DW. Evidence of multiple extracellular phospholipase activities of Aspergillus fumigatus. Infect Immun 1996; 64: 751-755.
Noodeh, AD. phospholipases of Aspergillus fumigatus.PhD thesis, University of Manchester. 2007.
Jepson M, Howells A, Bullifent HL, Bolgiano B, Crane D, Miller J, et al . Differences in the carboxy- terminal (Putative phospholipid binding) domains of Clostridium perfringens and Clostridium bifermentans phospholipases C influence the hemolytic and lethal properties of these enzymes. Infect Immun 1999; 67:3297-3301.
Sambrook J, Russel, DW. Quantitaive PCR. In: Sambrook J, Russel, DW, Molecular Cloning laboratory Manual. Cold Spring Harbor Laboratory Press; 2001:8.94.
Brisson M, Tan L, Park R, Hamby K. Identification of Nonspecific Products Using Melt-Curve Analysis on the iCycler iQ Detection System. Bio RAD tech note 2000: 1-6.
Schmiel DH, Miller VL. Bacterial phospholipases and pathogenesis. Microbes Infect; 1999. 1: 1103-1112.
de Vries RP, Visser J. Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. Microbiol Mol Biol Rev 2001; 65: 497-522.
Tonukari NJ. Enzymes and fungal virulence. J App Sci Env Manag 2003; 7: 5-8.
Birch M, Denning DW, Robson GD. Comparison of extracellular phospholipase activities in clinical and environmental Aspergillus fumigatus isolates. Med Mycol 2004; 42: 81-86.
Krawiec S, Riley, M. Organization of the bacterial chromosome. Microbiol Rev 1990; 54: 502-539.
Lawrence JG, Selfish operons and speciation by gene transfer. Trends Microbiol 1997; 5: 355-359.
Martin W. Mosaic bacterial chromosomes: a challenge en route to a tree of genomes. Bioessays 1999; 21: 99-104.
Flint HJ. Molecular genetics of obligate anaerobes from the rumen. FEMS Microbiol Lett 1994; 121: 259-267.
Rosewich UL, Kistler HC. Role of horizontal gene transfer in the evolution of fungi. Annu Rev Phytopathol 2000; 38: 325-363.
Garcia-Vallve S, Romeu A, Palau J. Horizontal gene transfer of glycosyl hydrolases of the rumen fungi. Mol Biol Evol 2000; 17: 352-361.
Temporini E, VanEtten H. An analysis of the phylogenetic distribution of the pea pathogenicity genes of Nectria haematococca MPVI supports the hypothesis of their origin by horizontal transfer and uncovers a potentially new pathogen of garden pea: Neocosmospora boniensis. Curr Genet 2004; 46: 29-36.
Wenzl P, Wong L, Kwang-won K, Jefferson R. A functional screen identifies lateral transfer of beta- glucuronidase (gus) from bacteria to fungi. Mol Biol Evol 2005; 22: 308-316.
Morgenroth K. The Surfactant System of the Lungs.Berlin: Walter de Gruyter; 1988.
Peberdy J. Protein secretion in filamentous fungi-trying to understand a highly productive black box. Trends Biotechnol 1994; 12: 50-57.
Pakula T, Salonen K, Uusitalo J, Penttila M. The effect of specific growth rate on protein synthesis and secretion in the filamentous fungus Trichoderma reesei. Microbiol 2005; 151: 135-143.
| Files | ||
| Issue | Vol 6 No 2 (2014) | |
| Section | Articles | |
| Keywords | ||
| Aspergillus fumigatus gene expression phosphatidylcholine phospholipase C plc gene | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
