Two Plasmodium falciparum ribonucleotide reductase small subunits, PfR2 and PfR4, interact with each other and are components of the in vivo enzyme complex

    Authors

    V. Bracchi-Ricard; D. Moe;D. Chakrabarti

    Comments

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    Abbreviated Journal Title

    J. Mol. Biol.

    Keywords

    malaria; Plasmodium falciparum; ribonucleotide reductase; deoxyribonucletide; CELL-CYCLE; DNA-DAMAGE; PROTEIN; YEAST; LOCALIZATION; EXPRESSION; MALARIA; IDENTIFICATION; PURIFICATION; CHECKPOINT; Biochemistry & Molecular Biology

    Abstract

    Ribonucleotide reductase (RNR) is a tetrameric enzyme, composed of two large (R1) and two small (R2) subunits, which regulates the nucleotide balance in cells by controlling the rate-limiting step for deoxyribonucleotide synthesis. We have identified a second copy of the small subunit gene, termed PfR4, encoding a 324 amino acid residue polypeptide that shares only 25% identity with the previously identified PfR2 small subunit of Plasmodium falciparum. PfR4 expression is cell-cycle-regulated, and the profile of transcript and protein expression corresponds to that of PfR2. A 1.3 kb PfR4 5 '-flanking fragment contained a functional promoter activity. We have detected interaction between PfR2 and PfR4 by co-immunoprecipitation experiments. Indirect immunofluorescence analysis showed distinct localization of two small RNR subunits with some colocalization. The association of PfR1 large subunit with PfR4 was detected by GST pull-down assay. This interaction is reduced significantly when using a PfR4 truncated at the COOH terminus, suggesting the involvement of COOH-terminal residues in PfR4-PfR1 interaction. All three RNR subunits co-eluted on a Superose 12 size-exclusion column corresponding to fractions with a molecular mass of around 250 kDa. This suggests the existence of all three RNR subunits in Plasmodium in a native complex of alpha(2)beta beta ' configuration. (c) 2005 Elsevier Ltd. All rights reserved.

    Journal Title

    Journal of Molecular Biology

    Volume

    347

    Issue/Number

    4

    Publication Date

    1-1-2005

    Document Type

    Article

    Language

    English

    First Page

    749

    Last Page

    758

    WOS Identifier

    WOS:000227959200006

    ISSN

    0022-2836

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