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Self-association of histones H1 from calf thymus and from sperm of the marine worm Chaetopterus variopedatus was studied on native and glutaraldehyde cross-linked molecules by PAGE and by salt-induced turbidity measurements. Multiple polymers were generated by native sperm histone H1-like after glutaraldehyde cross-linking while the same treatment on its lysine- or arginine-modified derivatives and on somatic histone H1 failed to induce polymerization. This result suggests the relevance of arginine content in the formation of histone H1-like polymers particularly because Chaetopterus variopedatus and calf thymus histones H1 have similar content of lysine but different K/R ratio (2 and 15, respectively). Salt-induced turbidity experiments confirmed the high tendency of sperm histone H1-like to form oligomers, particularly in the presence of phosphate ions. Native PAGE analysis in the presence of phosphate supported this hypothesis. The reported results suggest that phosphate ions connecting lysine and arginine side chain groups contribute to the interaction of sperm histone H1-like with DNA in chromatin and mplay a key role in organization and stabilization of the chromatin higher order structures.
As continuation of investigations in to the mechanism of the role of the H1 histone, which is a crucial protein component chromosomes of all eukaryotes, transgenic tobacco plants with different levels of the H1 histone variants were examined. Tobacco has six sequential variants of the H1 histone: two major ones (H1A and H1B), constituting ca. 90% of all H1, and four minor ones (H1C, H1D, H1E and H1F), occurring in very small quantities. The following groups of plants were examined: K - control group with a full set of histone variants; -AB -with the A and B variants removed; -ABCD - with the A, B, C and D variants removed; and -CD - with the C and D variants removed. The analysis of microsporogenesis in those plants, based on preparations squeezed in acetoorcein, revealed the asynchronous course of meiosis in -AB and -ABCD plants, occurrence of chromosomal aberration, and, consequently, the formation of sterile pollen grains (accordingly: 84,4% and 81,4%). In -CD plants, the percentage of aberration and sterile pollen grains was similar to the control material. Electron microscope observations of microsporogenesis showed ultrastructural changes. In -AB and -ABCD plants, a major portion of the pollen grains were degraded. The smallest number of degraded pollen grains, in comparison with the control, was found in the -CD group.
The role of histone H1 in the regulation of flower growth and development and in microsporogenesis was investigated in previously obtained transgenic tobacco plants expressing the antisense histone H1 cDNA. These plants exhibited a characteristic male-sterility phenotype caused by altered proportions of the native histone H1 in chromatin. The type and frequency of chromosomal aberrations during male meiosis as well as the distortions affecting tetrad formation in transgenic plants are reported.
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