Association of Bdf1 and Swc7 is dependent on the N terminus of Swr1.
A, SDS-PAGE and Western blotting analysis of Bdf1 and Swc7 in partial
SWR1 complexes purified from wild-type and mutant swr1 strains as
indicated. PVDF membrane strips corresponding to subunit(s) were probed with
antibodies against Bdf1 and Swc7. The schematic diagram of Swr1 indicates
subunit interaction domains of Swr1 and fragments used in this study:
N1 (residues 1–210), N2 (residues 278–681),
Insert (residues 1002–1221), Swr1 (residues
1–681). SWR1(ΔN1), SWR1(ΔN2), and SWR1(ΔInsert) are
mutant SWR1 complexes isolated from cells lacking the N1, N2, and Insert
regions of the Swr1 ATPase subunit, respectively
(32). B, SDS-PAGE and
Western blotting analysis of Swr1, Bdf1, Arp4, Swc4, Yaf9, and Swc7 in SWR1
complexes isolated from wild-type, bdf1Δ and
swc7Δ strains. PVDF membranes were probed with anti-Flag (to
reveal Swr1), and antibodies against individual SWR1 subunits as indicated.
C, SDS-PAGE (4–20% gel) and Western blotting analysis of Yaf9,
Bdf1, and Swc7 in SWR1 complexes isolated from wild-type and
yaf9Δ cells. PVDF membranes were probed with antibodies against
Yaf9, Bdf1, and Swc7, respectively.