Figure 5.

FACT helps to restore cac1∆ rtt106∆-mediated chromatin assembly defects. (A) Plasmid topoisomer distribution of the 2µ plasmid in wild type, hir1∆, cac1∆ rtt106∆ and cac1∆ rtt106 hir1∆ cells synchronized in G1 and released into fresh medium for different times. (B) Plasmid topoisomer distribution of the 2µ plasmid in wild type, spt16-G132D, cac1∆ rtt106∆ and cac1∆ rtt106∆ spt16-G132D cells synchronized in G1 at 26 °C and released into fresh medium at 31 °C; in S phase, cells were collected, washed to eliminate the pronase and released into pre-heated medium with α-factor till the following G1. (C) Plasmid topoisomer distribution of the 2µ plasmid in wild type, spt16-m, cac1∆ rtt106∆ and cac1∆ rtt106∆ spt16-m cells synchronized in G1 and released into fresh medium; in S phase, cells were collected, washed to eliminate the pronase and released into pre-heated medium with α-factor till the following G1. The experiment was repeated twice with similar results. (A–C) Cell cycle progression and topoisomer profiles are shown. r and SC(−) indicate relaxed and negative supercoiling, respectively. Cropped images show only relaxed and negatively supercoiled topoisomers. Original gels are presented in Fig. S5. The experiments were repeated at least twice with similar results.