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 Citations (Scopus)

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 - 1992

Fingerprint

Adenosylhomocysteinase
Antiparasitic Agents
Leishmania donovani
Organism Cloning
Drug Therapy
Genes
S-Adenosylmethionine
Parasitic Diseases
Proteins
Genomic Library
Homeobox Genes
DNA
Adenine
Enzymes
Southern Blotting
Eukaryota
Drosophila melanogaster
Northern Blotting
Open Reading Frames
Amino Acid Sequence

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Parasitology

Cite this

Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy. / Henderson, Debbie M.; Hanson, Sheri; Allen, Thomas; Wilson, Keith; Coulter-Karis, Donna E.; Greenberg, Michael L.; Hershfield, Michael S.; Ullman, Buddy.

In: Molecular and Biochemical Parasitology, Vol. 53, No. 1-2, 1992, p. 169-183.

Research output: Contribution to journalArticle

Henderson, DM, Hanson, S, Allen, T, Wilson, K, Coulter-Karis, DE, Greenberg, ML, Hershfield, MS & Ullman, B 1992, 'Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy', Molecular and Biochemical Parasitology, vol. 53, no. 1-2, pp. 169-183. https://doi.org/10.1016/0166-6851(92)90019-G
Henderson, Debbie M. ; Hanson, Sheri ; Allen, Thomas ; Wilson, Keith ; Coulter-Karis, Donna E. ; Greenberg, Michael L. ; Hershfield, Michael S. ; Ullman, Buddy. / Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy. In: Molecular and Biochemical Parasitology. 1992 ; Vol. 53, No. 1-2. pp. 169-183.
@article{3a13f90c416048ac9cfeeeb2cf27ce55,
title = "Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy",
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.",
keywords = "Chemotherapy, Leishmania donovani, Methylation, S-Adenosylhomocysteine hydrolase, S-Adenosylmethionine",
author = "Henderson, {Debbie M.} and Sheri Hanson and Thomas Allen and Keith Wilson and Coulter-Karis, {Donna E.} and Greenberg, {Michael L.} and Hershfield, {Michael S.} and Buddy Ullman",
year = "1992",
doi = "10.1016/0166-6851(92)90019-G",
language = "English (US)",
volume = "53",
pages = "169--183",
journal = "Molecular and Biochemical Parasitology",
issn = "0166-6851",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Cloning of the gene encoding Leishmania donovani S-adenosylhomocysteine hydrolase, a potential target for antiparasitic chemotherapy

AU - Henderson, Debbie M.

AU - Hanson, Sheri

AU - Allen, Thomas

AU - Wilson, Keith

AU - Coulter-Karis, Donna E.

AU - Greenberg, Michael L.

AU - Hershfield, Michael S.

AU - Ullman, Buddy

PY - 1992

Y1 - 1992

N2 - 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.

AB - 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.

KW - Chemotherapy

KW - Leishmania donovani

KW - Methylation

KW - S-Adenosylhomocysteine hydrolase

KW - S-Adenosylmethionine

UR - http://www.scopus.com/inward/record.url?scp=0026762577&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026762577&partnerID=8YFLogxK

U2 - 10.1016/0166-6851(92)90019-G

DO - 10.1016/0166-6851(92)90019-G

M3 - Article

VL - 53

SP - 169

EP - 183

JO - Molecular and Biochemical Parasitology

JF - Molecular and Biochemical Parasitology

SN - 0166-6851

IS - 1-2

ER -