title: Cloning and partial characterization of the proteasome S4 ATPase from Plasmodium falciparum.
abstract: Certad, G., Abrahem, A., and Georges, E. 1999. Cloning and Partial characterization of the proteasome S4 ATPase from Plasmodium falciparum. Experimental Parasitology 93, 123-131. The ATP-ubiquitin-proteasome pathway mediates the nonlysosomal degradation of cytosolic proteins in eukaryotic cells. The activities of this pathway have been shown to regulate cell growth and differentiation through modulation of regulatory proteins. The proteasome is a large complex consisting of two multisubunit structures, the 20S and 19S(PA700) or P28 complexes, that combine to form the 26S particles. In this study, we describe the cloning of a cDNA encoding the proteasome subunit 4 ATPase homologue from Plasmodium falciparum (PFS4). Analysis of the PFS4 cDNA sequence shows an open reading frame encoding a deduced protein of 455 amino acids. Moreover, comparison of PFS4 cDNA sequence to that of genomic fragments encoding PFS4 showed identical sequences with no detectable introns. Database searches revealed a high sequence identity to those of rice, yeast, mouse, Drosophila, and human S4 ATPases. However, PFS4 contains two unique inserts of nine and seven amino acid residues in the N-terminal domain. Interestingly, only the rice S4 contains the latter (seven amino acids) insert with four identical amino acids. In vitro expression of the full-length cDNA encoding the PFS4, using a transcription-translation-coupled reticulocyte lysate, shows a 50-kDa [(35)S]methionine-labeled protein which was immunoprecipitated with PFS4 anti-peptide antiserum. Southern blot analysis of genomic DNA digests shows a single gene copy of PFS4 in P. falciparum. Of interest was the effect of the proteasome-specific natural product, lactacystin, on the growth of the parasite, with IC(50) values of 0.6-0.92 microM. The latter IC(50) values of lactacystin for different clones of P. falciparum are comparable to those obtained for mammalian cell lines (0.65 microM), suggesting the presence of a conserved proteasome complex. Moreover, lactacystin was equally toxic to drug-sensitive and resistant parasites.
authors.affiliation: Institute of Parasitology, McGill University, Ste-Anne de Bellevue, Quebec, Canada.
authors.forename: G
authors.initials: G
authors.lastname: Certad
authors.forename: A
authors.initials: A
authors.lastname: Abrahem
authors.forename: E
authors.initials: E
authors.lastname: Georges
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000235:genetics; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D018076:DNA, Complementary / Q000737:chemistry; D016054:DNA, Protozoan / Q000737:chemistry; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010963:Plasmodium falciparum / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D016053:RNA, Protozoan / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D015853:Cysteine Proteinase Inhibitors; D018076:DNA, Complementary; D016054:DNA, Protozoan; D016053:RNA, Protozoan; C067713:lactacystin; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C109306:S4 ATPase, proteasome; D000111:Acetylcysteine