Remodeling of chromatin during male pronucleus formation in the sea urchin Tetrapygus niger.

Abstract

After fertilization the cone shaped sperm nucleus is transformed into a spheroidal male pronucleus which fuses with the female pronucleus reestablishing the diploid genome. These morphological changes are correlated with biochemical transitions of sperm-specific chromosomal proteins. To obtain information on the rearrangements of chromosomal proteins after fertilization, we have followed sperm-specific nonhistones (Sp NHCP) and the sperm-specific histones (SpH) in zygotes harvested at different times post-insemination (p.i.). The acquisition of proteins synthesized de novo was determined during the time of male pronucleus formation, estimated to occur until 30 min p.i. The results obtained may be summarized as follows: 1. The fate of Sp NHCP, followed by Western blot analysis with polyclonal antibodies obtained in rabbits against the whole complement of Sp NHCP, indicates that the majority of Sp NHCP are lost shortly p.i., except two proteins of 58 Kd and 61 Kd that were not distinguishable from eggs NHCP because a crossreaction was obtained. Acidic proteins synthesized de novo were bound to chromatin between 3 and 30 min p.i., and the majority of these proteins comigrated with egg NHCP in SDS/PAGE. 2. The histones from sperm differ from their counterparts found in unfertilized eggs, both in their amino acid composition and their microheterogeneity in bidimensional gels. Sperm contain the five major SpH whereas eggs have seven major histone variants, distinct from each other, with an electrophoretic behaviour consistent with histones but a different amino acid composition. The total complement of histone variants found in zygotes collected at amphimixis is identical with that found in unfertilized eggs, suggesting that SpH should be lost before that time.(ABSTRACT TRUNCATED AT 250 WORDS)