Nucleosome organization during germ cell development in the sea cucumber Holothuria tubulosa.

Abstract

Conformational changes that occur in chromatin from developing germ cells of the echinoderm Holothuria tubulosa have been probed with micrococcal nuclease. The results indicate that the extent of DNA degradation to acid-soluble nucleotides is highest in chromatin at the early stages of gonad growth, being drastically subdued in the mature sperm cell. Production of nucleosomal particles also varies with development, involving at least 70% of the chromatin at the final stage of maturation, whereas in immature germ cells it remains much lower. In contrast, electrophoretic analysis for DNA size has shown that the average nucleosome repeat length, about 227 base pairs, does not change throughout the maturation process. However, kinetics of the enzyme reaction have revealed that, although brief digestion of chromatin from both immature gonads and sperm yields comparable series of higher oligomers, extensive digest patterns differ widely. Sperm chromatin, highly protected, releases a 275 base pair intermediate fragment, wholly absent in immature gonads. The 145 base pair core released in both chromatins is not further digested in sperm. In comparison to sperm chromatin, that of immature germ cells is much more susceptible to fragmentation, yielding the usual set of smaller subnucleosomal fragments. These data suggest the induction of differential accessibilities of chromatin DNA with maturation, which is not accompanied by displacement of the histone complement. The histone variants present in this species may well be instrumental in the process.