Genetic nomenclature for Trypanosoma and Leishmania

Clayton, Christine; Adams, Mark; Almeida, Renata; Baltz, Théo; Barrett, Mike; Bastien, Patrick; Belli, Sabina I.; Beverley, Stephen M.; Biteau, Nicolas; Blackwell, Jenefer M.; Blaineau, Christine; Boshart, Michael; Bringaud, Frédéric; Cross, George A.M.; Cruz, Ângela K.; Degrave, Win; Donelson, John E.; El-Sayed, Najib M.; Fu, G; Ersfeld, Klaus; Gibson, Wendy; Gull, Keith; Ivens, Alasdair; Kelly, John M.; Vanhamme, Luc

doi:10.1016/s0166-6851(98)00115-7

Cited by 83 publications

(45 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The corresponding plasmid was obtained (courtesy Philip Majiwa, ILRI) and sequenced. The cDNA fragment contained the 3Ј-part of a cDNA that unambiguously represented a phosphodiesterase gene, termed TbPDE2A according to the recently proposed rules for the nomenclature of trypanosomatid genes (43). Southern blot analysis of genomic DNA demonstrated that TbPDE2A is not a single gene but rather a member of a small gene family ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Characterization of TbPDE2A, a Novel Cyclic Nucleotide-specific Phosphodiesterase from the Protozoan Parasite Trypanosoma brucei

Zoraghi¹,

Kunz

Gong³

et al. 2001

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

This study reports the identification and characterization of a cAMP-specific phosphodiesterase from the parasitic hemoflagellate Trypanosoma brucei. TbPDE2A is a class I phosphodiesterase. Its catalytic domain exhibits 30 -40% sequence identity with those of all 11 mammalian phosphodiesterase (PDE) families, as well as with PDE2 from Saccharomyces cerevisiae, dunce from Drosophila melanogaster, and regA from Dictyostelium discoideum. The overall structure of TbPDE2A resembles that of human PDE11A in that its N-terminal region contains a single GAF domain. This domain is very similar to those of the mammalian PDE2, -5, -6, -10, and -11, where it constitutes a potential cGMP binding site. TbPDE2A can be expressed in S. cerevisiae, and it complements an S. cerevisiae PDE deletion strain. Recombinant TbPDE2A is specific for cAMP, with a K m of ϳ2 M. It is entirely resistant to the nonselective PDE inhibitor 3-isobutyl-1-methylxanthine, but it is sensitive to trequinsin, dipyridamole, sildenafil, and ethaverine with IC 50 values of 5.4, 5.9, 9.4, and 14.2 M, respectively. All four compounds inhibit proliferation of bloodstream form trypanosomes in culture, indicating that TbPDE2A is an essential enzyme.

show abstract

Section: Resultsmentioning

confidence: 99%

Characterization of TbPDE2A, a Novel Cyclic Nucleotide-specific Phosphodiesterase from the Protozoan Parasite Trypanosoma brucei

Zoraghi¹,

Kunz

Gong³

et al. 2001

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Transfected parasites were then selected in 20 g/ml G418 in DME-L-CS medium supplemented with 200 M putrescine and 200 M ornithine. Transfectant lines harboring episomes encompassing the wild type and COOH-terminal deletion constructs were designated ⌬arg[pGFP-arginase] and ⌬arg[pGFP-arg⌬skl], respectively, according to the generally accepted genetic nomenclature for Leishmania (41).…”

Section: Methodsmentioning

confidence: 99%

Arginase Plays a Pivotal Role in Polyamine Precursor Metabolism in Leishmania

Roberts

Tancer

Polinsky

et al. 2004

Journal of Biological Chemistry

159

193

View full text Add to dashboard Cite

The polyamine pathway of protozoan parasites has been successfully targeted in anti-parasitic therapies and is significantly different from that of the mammalian host. To gain knowledge into the metabolic routes by which parasites synthesize polyamines and their precursors, the arginase gene was cloned from Leishmania mexicana, and ⌬arg null mutants were created by double targeted gene replacement and characterized. The ARG sequence exhibited significant homology to ARG proteins from other organisms and predicted a peroxisomal targeting signal (PTS-1) that steers proteins to the glycosome, an organelle unique to Leishmania and related parasites. ARG was subsequently demonstrated to be present in the glycosome, whereas the polyamine biosynthetic enzymes, in contrast, were shown to be cytosolic. The ⌬arg knockouts expressed no ARG activity, lacked an intracellular ornithine pool, and were auxotrophic for ornithine or polyamines. The ability of the ⌬arg null mutants to proliferate could be restored by pharmacological supplementation, either with low putrescine or high ornithine or spermidine concentrations, or by complementation with an arginase episome. Transfection of an arg construct lacking the PTS-1 directed the synthesis of an arg that mislocalized to the cytosol and notably also complemented the genetic lesion and restored polyamine prototrophy to the ⌬arg parasites. This molecular, biochemical, and genetic dissection of ARG function in L. mexicana promastigotes establishes: (i) that the enzyme is essential for parasite viability; (ii) that Leishmania, unlike mammalian cells, expresses only one ARG activity; (iii) that the sole vital function of ARG is to provide polyamine precursors for the parasite; and (iv) that ARG is present in the glycosome, but this subcellular milieu is not essential for its role in polyamine biosynthesis.

show abstract

“…We have used the standard trypanosomatid nomenclature for genes and proteins, whereby genes are upper case and italicized, proteins upper case. 18 Only around half of the yeast proteins yield Proteins with broken underlining are specific for the Cvt transport process, those continuously underlined for the autophagy pathways in yeast; proteins not underlined are shared by both processes. For proteins shown in grey, orthologues have been confidently identified in at least one of the trypanosomatid databases.…”

Section: Introductionmentioning

confidence: 99%