Effect of the C-terminal deletion of Xrs2 on DNA sensitivity and the
complex formation. (A,B) Sensitivity to phleomycin (A) and
UV light (B). Serial dilutions of cultures were spotted on YEPD media
without or with phleomycin. Cultures spotted on YEPD media were irradiated
with UV light. All of the strains contain the sml1Δ mutation
that suppresses the lethality of mec1 mutants
(Zhao et al. 1998). Strains
used were wild-type (KSC1560), xrs2Δ (KSC1562),
xrs2-11 (KSC1563), tel1Δ (KSC1661), mec1-81
(KSC1662), mec1-81 tel1Δ (KSC1564), mec1-81 xrs2-11
(KSC1565), and mec1-81 tel1Δ xrs2-11 (KSC1566).
(C) Interaction between Tel1and Xrs2. Cells were untreated (–)
or treated with 50 μg/mL phleomycin for 1h (+). Extracts were prepared from
cells and immunoprecipitated with anti-HA antibodies. Immunoprecipitates (IP)
and whole extracts were subjected to immunoblotting analysis. Strains used
were XRS2-myc (KSC1904), xrs2-11-myc (KSC1905),
TEL1-HA (KSC1785), TEL1-HA XRS2-myc (KSC1906), and
TEL1-HA xrs2-11-myc (KSC1907). (D) Interaction of Xrs2 with
Mre11 and Rad50. Extracts were prepared from XRS2-HA (KSC1744),
xrs2-11-HA (KSC1869), or untagged cells (KSC1516), and
immunoprecipitated with anti-HA antibodies. IPs and whole extracts were
subjected to immunoblotting analysis. Phosphatase treatment diminished the
slower migrating forms of Xrs2-HA and Xrs2-11-HA (data not shown), suggesting
that Xrs2 and Xrs2-11 are phosphorylated proteins.