TARGETED DISRUPTION OF 01H1 ENCODING A PARTICULAR H1 HISTONE VARIANT CAUSES CHANGES IN PROTEIN-PATTERNS IN THE DT40 CHICKEN B-CELL LINE

Six members of the chicken H1 gene family, all of which are located in two major histone gene clusters, have been shown to encode six different protein variants. The intracellular mRNA level from one of them, 01H1, encoding the 01H1 variant composed of 218 amino acid residues, constitutes 9.9% of the total H1 mRNA in the DT40 chicken B cell line. To study the specific role of this particular H1 variant, besides its well-known functions as a linker in chromatin maintenance and as a general repressor of transcription, we used targeted integration to construct heterozygous and homozygous DT40 mutants with disruption of one and two 01H1 alleles, respectively. Analyses of the stable transfectants showed that the growth rate of DT40 was unchanged in the absence of two 01H1 alleles. Moreover, the remaining H1 genes were shown to be expressed more in these mutants than in the wild-type cell lines. Two-dimensional polyacrylamide gel electrophoresis showed that within an almost constant background in the homozygous mutants several cellular proteins newly appeared or increased, while some other proteins disappeared or decreased quantitatively; These variable proteins all differed from those that varied in DT40 mutants deprived of one of the eight thicken H2B genes, H2B-V, encoding a particular H2B variant. These results suggest that the 01H1 variant is involved in the regulation of expression of genes that encode the proteins that vary in 01H1-deleted mutants of DT40 cells. (C) 1995 Academic Press Limited