Figure 4.—

Dominant-negative mutant Ltv1 is incorporated into subunits and causes a 40S deficit. LY134 (wild-type) cells were cotransformed with empty vector (pUG23), Ld47 (P_MET–LTV1–GFP), or Ld57 (P_MET–ltv1_Δ169-176–GFP). Extracts were prepared and sedimented through sucrose density gradients as described in materials and methods. The presence of wild-type or mutant Ltv1–GFP and Rps3 in individual fractions was determined by Western blotting using anti-GFP and anti-Rps3 antibodies, respectively. Fractions containing 80S and deeper fractions were combined as indicated.