Skip to main content
. 2011 Jul 29;6(7):e22209. doi: 10.1371/journal.pone.0022209

Figure 6. htb1-T122E dominantly impairs telomeric chromatin formation.

Figure 6

(A–B) The effects of H2B C-terminal mutations on association of SIR proteins in telomeric regions. Strains derived from Y131 expressing HTB1, htb1-T122A and T122E were analyzed by chromatin immunoprecipitation using α-Sir2 Ab (A) or α-myc Ab to detect myc-tagged Sir3 (B). (C–D) The effect of T122E on chromatin mobility is SIR complex independent. Distribution of chromatin from a strain derived from UCC6389 expressing htb1-T122E and strains derived from UCC6391 (sir4Δ) expressing HTB1 (or htb1-T122E) were analyzed by 20%–40% sucrose gradient ultracentrifugation. The relative positions of the fragments containing sub-telomeric (sub-TEL), or euchromatin region (euC), were shown in (C) or (D), respectively. (E–F) The dominance of htb1 mutants was analyzed by telomere silencing assay. Plasmids carrying HTB1 or htb1 mutants were transformed into strain UCC6389 expressing HTB1 (E), or plasmids carrying HTB1 were transformed into the strain UCC6389 expressing HTB1 or htb1 mutants (F). Overnight cultures of the indicated cells were 5-fold serial diluted on the indicated plates.