Figure 9.

Formation of an I1 complex in the rescued ida2 strain. Dynein extracts from wild type and the rescued ida2-6 strain, D11, were subjected to sucrose density gradient centrifugation. The resulting fractions were run on duplicate 5–15% polyacrylamide gradient gels, and the gels were either silver stained or transferred to Western blots. (A) Gel of wild-type fractions sedimenting in the 19S region. The 110-, 138-, and 140-kDa ICs of the I1 complex (indicated by the solid circles) cosediment in fractions 6 and 7. (B) Gel of the rescued ida2-6 strain, D11. The I1 ICs cosediment at 16S in fractions 9 and 10. A novel polypeptide migrating at ∼113 kDa in fractions 9 and 10 is indicated by a diamond. This polypeptide is not present in the wild-type gradient. (C) Blots of the wild-type fractions shown in A were probed with antibodies against the 1α Dhc (top panel), IC140 (second panel), 1β Dhc (third panel), and Tctex1 LC (bottom panel). (D) Blots of the rescued ida2-6 fractions shown in B were probed with I1 complex antibodies as described in C. Note the presence of the 1β Dhc fragment at ∼113 kDa.