Problem:
Before nuclear pore complexes were well understood, it was unclear whether nuclear proteins diffused passively into the nucleus and accumulated there by binding to residents of the nucleus such as chromosomes, or were actively transported and accumulated regardless of their affinity for nuclear components.
A classic experiment that addressed this problem used several forms of radioactive nucleoplasmin, which is a large pentameric protein involved in chromatin assembly. In this experiment, either the intact protein or the nucleoplasmin heads, tails, or heads with a single tail were injected into the cytoplasm or into the nucleus of frog oocytes. All forms of nucleoplasmin, except heads, accumulated in the nucleus when injected in the cytoplasm, and all forms were retained in the nucleus when injected there.
Required:
Question 1: What portion of the nucleoplasmin molecule is responsible for localization in the nucleus?
Question 2: How do these experiments distinguish between active transport, in which a nuclear localization signal triggers transport by the nuclear pore complex, and passive diffusion, in which a binding site for a nuclear component allows accumulation in the nucleus?
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