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. 2012 Jan 23;7(1):e30622. doi: 10.1371/journal.pone.0030622

Figure 5. Skp1 together with Fbh1 resolves meiotic recombination intermediates.

Figure 5

(A) Temperature sensitivity of the skp1-a7 mutant was partially suppressed by overexpression of Fbh1 from the plasmid vector pREP1. Ten-fold serial dilution of the indicated strains was spotted on minimal media with or without thiamine and then incubated at 25°C and 36°C. Overexpression is induced on the medium without thiamine (−thiamine). (B) Growth defects of skp1-a7 and fbh1Δ mutants were suppressed by rad3Δ, removal of the Rad3-dependent DNA damage checkpoint. (C, D) Frequency of the normal and bent spindle in mitosis (C) and the aberrant ‘crossing’ spindles in MII (D) in the indicated strains. n≥30. (E) Schematic diagram for Fbh1 WT and Fbh1-D485N (the helicase-dead form), Fbh1-L14A P15A and Fbh1-P15A L26A (the F-box-deficient form) mutant proteins. (F) Sensitivity to 5 µM CPT (camptothecin) was tested for the indicated strains. (G) fbh1-D485N and fbh1-P15A L26A mutants exhibited the bent-spindle phenotype, whereas fbh1-L14A P15A did not. GFP-Atb2 in MI of the indicated strains is shown. Bars, 5 µm.