Skip to main content
Figure 3 | Malaria Journal

Figure 3

From: The dimerization domain of Pf CENP-C is required for its functions as a centromere protein in human malaria parasite Plasmodium falciparum

Figure 3

Functional complementation of Saccharomyces cerevisiae MIF2p by the Pf CENP-C motif and Pf CENP-C dimerization domain. (A and B, i-iii) Growth and ten-fold serial dilution assays with mif2-3 (i and ii) and mif2-2 (iii) TS mutants expressing MIF2p, empty vector, various Pf CENP-C domains: NTD, motif, and DD and the parent TS strains at 23 and 37°C, respectively. (i) At 23°C, the mif2-3 TS expressing abovementioned constructs display normal growth. (ii and iii) At 37°C, TS mutants, expressing Pf CENP-C-NTD, vector alone and the parent TS cells alone fail to grow, while MIF2p shows complete growth restoration. The mif2-3 TS mutants expressing Pf CENP-C motif and DD show partial and complete growth rescue, respectively while in mif2-2 TS cells, both Pf CENP-C motif as well as DD show partial growth rescue. The red boxes highlight the comparative growth of various Pf CENP-C domains at 37°C. (C) Western blot analysis showing the expression of GFP tagged Pf CENP-C motif (~29.68 kDa) and DD (~38.16 kDa) in mif2-3 TS mutants at 23 (lanes 4 and 5) and 37°C (lanes 1 and 2). (D) Confocal images of GFP tagged Pf CENP-C motif and the DD (green) showing their mislocalization (red arrows) to the non-centromeric sites both at 23 and 37°C. (E) Confocal images represent the phenotypes of mif2-3 and mif2-2 TS mutants carrying various constructs at 23 and 37°C, respectively. At 23°C, all the transformants show normal bud morphology with equal nuclear (blue) segregation. At 37°C, like parent TS mutants, Pf CENP-C-NTD and empty vector show aberrant bud morphology with fragmented nuclei. The Pf CENP-C motif expressing mif2-3 and mif2-2 TS mutants shows chromosome mis-segregation (black arrows) and proper chromosome segregation, respectively. However, the Pf CENP-C-DD expression in both the TS mutants restores the normal chromosome segregation. MIF2p expressing cells also exhibit normal phenotypes at 37°C. N-terminal region-NTD, dimerization domain-DD.

Back to article page