Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy

Debbie M. Henderson, Sheri Hanson, Thomas Allen, Keith Wilson, Donna E. Coulter-Karis, Michael L. Greenberg, Michael S. Hershfield, Buddy Ullman

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

A full-length gene encoding the S-adenosylhomocysteine hydrolase (AdoHcyase) enzyme has been isolated from a genomic library of Leishmania donovani DNA in λ GEM-11 by cross-hybridization to the full-length human AdoHcyase cDNA. The nucleotide sequence of the SalI fragment contained a single open reading frame that encoded a polypeptide of 438 amino acids (47 712 Da). After maximum gap alignment, the predicted amino acid sequence of the leishmanial Ado Hcyase was 70-73% identical to AdoHCyases from higher eukaryotes. In addition, a data base search revealed that the primary structure of all AdoHcyase proteins was highly homologous to that of a protein encoded by a mRNA from Drosophila melanogaster that maps near the r element function of the Abd-b homeotic gene. In Northern blots, the SalI fragment hybridized to a 3.0-kb transcript that presumably encodes the parasite enzyme. Southern blot analysis of genomic DNA revealed that the AdoHcyase gene did not exist as a tandemly repeated array within the L. donovani genome. Moreover, monoclonal antibodies generated against human AdoHcyase recognized a leishmanial protein on immunoblots. Finally, the growth of L. donovani promastigotes could be arrested by micromolar concentrations of 3-deazaaristeromycin (C3Ari) and 9-(trans-2′, trans-3′-dihydroxycyclopentanyl)adenine, 2 known inhibitors of mammalian AdoHcyase. C3Ari also induced a substantial expansion of the intracellular pools of both AdoHcy and S-adenosylmethionine (AdoMet), as well as a significant diminution of the AdoMet/AdoHcy ratio. Thus, AdoHcyase may have therapeutic potential for the selective treatment of diseases of parasitic origin.

Original languageEnglish (US)
Pages (from-to)169-183
Number of pages15
JournalMolecular and Biochemical Parasitology
Volume53
Issue number1-2
DOIs
StatePublished - Jul 1992

Keywords

  • Chemotherapy
  • Leishmania donovani
  • Methylation
  • S-Adenosylhomocysteine hydrolase
  • S-Adenosylmethionine

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

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    Henderson, D. M., Hanson, S., Allen, T., Wilson, K., Coulter-Karis, D. E., Greenberg, M. L., Hershfield, M. S., & Ullman, B. (1992). Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy. Molecular and Biochemical Parasitology, 53(1-2), 169-183. https://doi.org/10.1016/0166-6851(92)90019-G