Fig. 5
Telomere protection function of Stn1-Ten1 is mediated by Tpz1 Lys242 SUMOylation and the SWSSS motif. a Detection of Tpz1 SUMOylation in wild-type and indicated mutant alleles of Tpz1. Denatured whole cell extracts from cells expressing endogenous wild-type Pmt3 only (lanes 1-4), cells ectopically over-expressing conjugatable His6-3HA-tagged Pmt3-gg (lanes 5-8) or non-conjugatable His6-3HA-tagged Pmt3-aa (lanes 9 and 10) from a pREP1-derived vector were processed for anti-myc western blot analysis. Additional western blots and their quantifications to determine changes in Tpz1 SUMOyltion levels are also shown in Supplementary Fig 3. b Southern blot analysis of telomere length for indicated mutant strains. For tpz1-K242R-AWAAA and tpz1-AWAAA pmt3-allR strains, hybridization signals for telomeric repeats were completely lost due to chromosome circularization. c Pulsed-field gel analysis of telomeric NotI-fragments. A schematic NotI restriction map of S. pombe chromosomes with telomeric NotI fragments (C, I, L, and M) from chromosomes I and II marked as black boxes is also shown
