Cryptosporidium parvum possesses a short-type replication protein A large subunit that differs from its host.
Replication protein A (RPA) consisting of three subunits is a eukaryotic single-stranded DNA (ssDNA)-binding protein involved in DNA replication, repair and recombination. We report here the identification and characterization of a RPA large subunit (CpRPA1) gene from the apicomplexan Cryptosporidium parvum. The CpRPA1 gene encodes a 53.9-kDa peptide that is remarkably smaller than that from other eukaryotes (i.e. approximately 70 kDa) and is actively expressed in both free sporozoites and parasite intracellular stages. This short-type RPA large subunit has also been characterized from one other protist, Crithidia fasciculata. Three distinct domains have been identified in the RPA large subunit of humans and yeasts: an N-terminal protein interaction domain, a central ssDNA-binding area, and a C-terminal subunit-interacting region. Sequence analysis reveals that the short-type RPA large subunit differs from that of other eukaryotes in that only the domains required for ssDNA binding and heterotrimer formation are present. It lacks the N-terminal domain necessary for the binding of proteins mainly involved in DNA repair and recombination. This major structural difference suggests that the mechanism for DNA repair and recombination in some protists differs from that of other eukaryotes. Since replication proteins play an essential role in the cell cycle, the fact that RPA proteins of C. parvum differ from those of its host suggests that RPA be explored as a potential chemotherapeutic target for controlling cryptosporidiosis and/or diseases caused by other apicomplexans.
Zhu
G
G
Wadsworth Center, New York State Department of Health, Albany 12201-2002, USA. zhug@wadsworth.org
Marchewka
M J
MJ
Keithly
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D016785:Cryptosporidium parvum / Q000737:chemistry* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D051716:Replication Protein A; D017386:Sequence Homology, Amino Acid
D004268:DNA-Binding Proteins; C497167:RPA1 protein, human; D051716:Replication Protein A