A. Summary of yeast two-hybrid interactions with various
Exo1 deletion constructs in prey vectors and their interactions with bait
vectors encoding Msh2 or Mlh1. The Exo1-F447A,F448A variant disrupts the Exo1
MIP box and the Mlh1 binding interaction. The Exo1Δ571–702 and
Exo1Δ571–635,Δ671–702 disrupt the Msh2 binding
sites, but not the Exo1-Mlh1 interaction. All experiments were independently
repeated a minimum of 4 times. B. Sequence alignment of the
Msh2-binding regions of S. cerevisiae Exo1 with closely related
fungal species in the Saccharomycotina are displayed so that residues identical
to S. cerevisiae are highlighted in green. C.
Sequence logos generated by Seq2Logo (ref) for alignments of SHIP box 1, SHIP
box 2, and both SHIP boxes based on an alignment of 291 fungal species; note
that the number of sequences for each SHIP box is lower than the total number of
fungal species analysed due to gene annotation errors and/or lack of
conservation of specific SHIP boxes. D. Yeast two-hybrid analysis
reveals that the Exo1-F582A,Y584A,Δ637–702 and
Exo1Δ571–635,F697A,Y699A constructs failed to interact with the
Msh2 bait (lack of growth on –Leu –Trp –His medium as
compared to growth on the control –Leu –Trp medium) but still
retained interaction with the Mlh1 bait. All experiments were independently
repeated a minimum of 4 times.