Kinetics of accumulation and depletion of soluble newly synthesized histone in the reciprocal regulation of histone and DNA synthesis.

Procedures are presented which permit the identification and analysis of cellular histone that is not bound to chromatin. This histone, called soluble histone, could be distinguished from that bound to chromatin by the state of H4 modification and the lack of H2A ubiquitination. Changes in the levels of newly synthesized soluble histone were analyzed with respect to the balance between histone and DNA synthesis in hamster ovary cells.

Comparative enzymatic degradation of H1 subfractions from Syrian hamster tissues.

An additional hydrolysis site recognized by thrombin on histone H1 molecules was found. Snakes venom proteases from Agkistrodon rhodostoma, Bothrops marajoensis and Bothrops moojeni were further used for the analysis of H1 histones. The presence of the main cleavage site on H1 histone molecules has been established.

Histones and their modifications.

Histones constitute the protein core around which DNA is coiled to form the basic structural unit of the chromosome known as the nucleosome. Because of the large amount of new histone needed during chromosome replication, the synthesis of histone and DNA is regulated in a complex manner. During RNA transcription and DNA replication, the basic nucleosomal structure as well as interactions between nucleosomes must be greatly altered to allow access to the appropriate enzymes and factors.

Distribution of the H1 histone subfractions in Syrian hamster chromatin fractions.

Chromatin from two Syrian hamster tissues: the Kirkman-Robbins hepatoma and the liver, has been separated into soluble (S) and insoluble (P) fractions. Both fractions contain the complete set of five main histones but differ in respect of H1 subfractions. The hepatoma chromatin is known to contain an unusual H1 subfraction, H1 slow [12, 13], probably identical with a similar subfraction present in hamster testes.

Occurrence of the low-mobility H1 histones subfraction in embryonic, differentiated, and neoplastic tissues of the Syrian hamster.

Electrophoretically slow H1 histone subfractions with mobilities identical to that of the subfraction found in the Kirkman-Robbins hamster hepatoma chromatin have been shown to be present in 12-day hamster embryos and in a sarcoma-type hamster tumor induced by SV40. No subfractions of such mobility were found in hamster liver, regenerating liver, thymus, spleen, and a fast-growing transplantable amelanotic hamster melanoma. A suggestion is made that some defective mechanisms of differentiation may affect the regulation of expression of the genes coding for the H1 histone subfractions.