An approximately half set of histone genes is enough for cell proliferation and a lack of several histone variants causes protein pattern changes in the DT40 chicken B cell line

Abstract

Of the 44 chicken histone genes, 39 are located in a major histone gene cluster of 110 kb, the others residing in four separate regions. The 42 sequenced genes encode six H1 variants, three H2A variants, four H2B variants, two H3 variants, and one histone H4. To clarify the influence on cell functions of simultaneous deletion of an approximately half set of the genes and some of the variants, we generated homozygous chicken DT40 mutants by disruption of two allelic segments of 57 kb, containing the 21 genes, using gene targeting techniques. Analyses with antisense RNA probes common or specific for gene families H1, H2A, H2B, H3 and H4 indicated that the remaining members of each of the gene families were expressed more in the mutants than in DT40 cells, resulting in maintenance of constant steady-state levels of mRNAs. Two-dimensional polyacrylamide gel electrophoresis showed that in the mutants several cellular proteins newly appeared or increased, and some other proteins disappeared or decreased quantitatively. These results demonstrate that all the histone gene families have the inherent ability to compensate for the disruption of a fair number of their own constituents. Furthermore, some of the histone variants are involved in regulation of the expression of putative genes that encode the proteins that varied in mutant DT40 cells, this participation is not compensated for by any residual variant of the same histone subtype(s).