Leishmania (Viannia) panamensis expresses a nuclease with molecular and biochemical features similar to the Endonuclease G of higher eukaryotes*
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Methods: The gene of the putative L. (V.) panamensis Endonuclease G was amplified, cloned, and sequenced. The recombinant protein was produced in a heterologous expression system and biochemical assays were run to determine its ion, temperature, and pH preferences.
Results: The L. (V.) panamensis rENDOG has biochemical features similar to those found in other trypanosomatids and higher eukaryotes. In addition, phylogenetic analysis revealed a possible evolutionary relationship with metazoan ENDOG.
Conclusions: L. (V.) panamensis has a gene that codifies an ENDOG homologous to those of higher organisms. This enzyme can be produced in Escherichia coli and is able to degrade covalently closed circular double-stranded DNA. It has a magnesium preference, can be inhibited by potassium, and is able to function within a wide temperature and pH range.
- Leishmania panamensis
- Endonuclease G
- Molecular characterization
- Recombinant proteins
- Refolding
- Phylogeny
Desjeux P. Leishmaniasis: current situation and newperspectives.Comp Immunol Microbiol Infect Dis. 2004; 27:305-18.2.Vélez ID, Gilchrist K, Arbeláez MP, Rojas CA, Puerta JA,Antunes CM, et al. Failure of a killed Leishmania amazonensisvaccine against American cutaneous leishmaniasis in Colom-bia.Trans R Soc Trop Med Hyg. 2005. 99: 593-8.3.Belkaid Y, Méndez S, Lira R, Kadambi N, MilonG, Sacks D.A natural model of Leishmania major infection reveals aprolonged «silent» phase of parasite amplification in the skinbefore the onset of lesion formation and immunity.JImmunol. 2000; 165: 969-77.4.Zandbergen V, Bollinger GA, Wenzel A, Kamhawi S, Voll R,Klinger M, et al. Leishmania disease development depends onthe presence of apoptotic promastigotes in the virulentinoculum.Proc Natl Acad Sci USA. 2006; 103: 13837-42.5.de Freitas JM, Moreira ME, Bonomo A, Bozza PT, AmaranteG, Pirmez C, et al. Apoptotic mimicry by an obligateintracellular parasite downregulates macrophage microbicidalactivity.Curr Biol. 2001; 11: 1870-3.6.Voll RE, Herrmann M, Roth EA, Stach C, Kalden JR,Girkontaite I. Immunosuppressive effects of apoptotic cells.Nature. 1997; 390: 350-1.7.Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY,Henson PM, Macrophages that have ingested apoptotic cellsin vitro inhibit proinflammatory cytokine production throughautocrine/paracrine mechanisms involving TGF-beta, PGE2,and PAF.J Clin Invest. 1998; 101: 890-8.8.Gannavaram S, Vedvyas C, Debrabant A, Conservation of thepro-apoptotic nuclease activity of endonuclease G in unicellulartrypanosomatid parasites.J Cell Sci. 2008; 121: 99-109.9.Rico E, Alzate JF, Arias AA, Moreno D, Clos J, Gago F, e t a l .Leishmania infantum expresses a mitochondrial nucleasehomologous to EndoG that migrates to the nucleus in responseto an apoptotic stimulus.Mol Biochem Parasitol. 2009; 163:28-38.10. Li LY, Luo X, Wang X. Endonuclease G is an apoptoticDNase when released from mitochondria.Nature. 2001; 412:95-9.11. Parrish J, Li L, Klotz K, Ledwich D, Wang X, Xue D,Mitochondrial endonuclease G is important for apoptosis in C.elegans.Nature. 2001; 412: 90-4.12. Low RL, Mitochondrial Endonuclease G function in apoptosisand mtDNA metabolism: a historical perspective.Mito-chondrion. 2003; 2: 225-36.13. Kuhlmann UC, Moore GR, James R, Kleanthous C, HemmingsAM. Structural parsimony in endonuclease active sites: shouldthe number of homing endonuclease families be redefined?FEBS Lett. 1999; 463: 1-2.14. Schafer P, Scholz SR, Gimadutdinow O, Cymerman IA,Bujnicki JM, Ruiz A, et al. Structural and functionalcharacterization of mitochondrial EndoG, a sugar non-specificnuclease which plays an important role during apoptosis.JMol Biol. 2004; 338: 217-28.15. Peacock CS, Seeger K, Harris D, Murphy L, Ruiz JC, QuailMA, e t a l. Comparative genomic analysis of three Leishmaniaspecies that cause diverse human disease.Nat Genet. 2007;39: 839-47.16. Marston FA. The purification of eukaryotic polypeptidessynthesized in Escherichia coli.Biochem J. 1986; 240: 1-12.17. Jerkovic B, Bolton PH. Magnesium increases the curvature ofduplex DNA that contains dA tracts.Biochemistry. 2001; 40:9406-11.18. Permyakov EA, Kretsinger RH. Cell signaling, beyondcytosolic calcium in eukaryotes.J Inorg Biochem. 2009; 103:77-86.19. Crawford DR, Abramova NE, Davies KJ. Oxidative stresscauses a general, calcium-dependent degradation of mito-chondrial polynucleotides.Free Radic Biol Med. 1998; 25:1106-11.20. Trump BF, Berezesky IK. Calcium-mediated cell injury andcell death.FASEB J. 1995; 9: 219-28.21. Bortner CD, Hughes FMJr, Cidlowski JA. A primary role forK+ and Na+ efflux in the activation of apoptosis.J Biol Chem.1997; 272: 32436-42.22. Hughes, FMJr, Bortner CD, Purdy GD, Cidlowski JA.Intracellular K+ suppresses the activation of apoptosis inlymphocytes.J Biol Chem. 1997; 272: 30567-76.23. Sen N, Das BB, Ganguly A, Mukherjee T, Bandyopadhyay S,Majumder HK. Camptothecin-induced imbalance inintracellular cation homeostasis regulates programmed celldeath in unicellular hemoflagellate Leishmania donovani.JBiol Chem. 2004; 279: 52366-75.24. Friedhoff P, Kolmes B, Gimadutdinow O, Wende W, KrauseKL, Pingoud A. Analysis of the mechanism of the Serratianuclease using site-directed mutagenesis.Nucleic Acids Res.1996; 24: 2632-9.
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