Figure 3.

Nmd3-bound 60S particles accumulate in the nucleus in an arx1Δ mutant. (A) Cultures of wild-type and arx1Δ cells carrying pAJ582 (Nmd3-GFP) were grown to midlog phase in selective media and the localization of Nmd3 was monitored by fluorescence microscopy. (B) Lysates were prepared from wild-type and arx1Δ cells and fractioned on 7–47% sucrose gradients by ultracentrifugation as described in Materials and Methods. Fractions were collected, and the absorbance at 254 nm was monitored continuously. Proteins were precipitated with trichloroacetic acid, separated by SDS-PAGE, transferred to nitrocellulose membrane, and immunoblotted for Nmd3 or Rpl8p using specific antibodies.