In vitro ribosomal ribonucleoprotein transport upon nuclear expansion

The interdependence of nuclear rRNA release and nuclear size is investigated in macronuclei isolated from Tetrahymena. Nuclei are induced to contract and to expand, without any structural disintegration of the nuclear envelope, by final Ca2+/Mg2+ (3:2) concentrations of 5 and 1.5 mM, respectively. Upon expansion, the average volume of nuclei increases from 600 +/- 42 to 811 +/- 76 micron3. Concomitantly, nuclei begin to release RNA following saturation kinetics. This RNA release stops immediately upon nuclear contraction. Similar to the in vivo situation, only advanced rRNA processing products are released in the form of ribosomal precursor particles, as identified in detail by polyacrylamide gel electrophoresis and rate zonal and isopycnic density gradient centrifugation. Three particle ty9es are released having average buoyant densities of 1.495, 1.470, and 1.532 g/cm3, exhibiting average sedimentation coefficients of 62, 62, and 35 S, and containing the immediate precursor to the 25S rRNA, 26S rRNA, and 17S rRNA, respectively. Tje rRNP release if ATP independent and noncoincident with the release of endogenous nuclear Pi, though it is Be2+ sensitive. Our data are compatible with the views that nuclear expansion is the prerequisite rather than the cause for the rRNP release and that nuclear pore complex associated ATPases play only, if at all, a minor role in nucleocytoplasmic exchange of rRNP.