Incorporation of various amino acids into non-histone chromatin protein fractions of spleen cells of mice immunized with IgG

Cell Biochemistry & Function

1985

Self Cite

Incorporation of [125I]IgG into spleen cells was studied in vivo and in vitro. In vivo, the antigen after uptake into the cytoplasm migrated into cell nuclei, where it was bound to chromatin up to the saturation level. One day after immunization the constant level of [125I]IgG was 1.3 X 10(12) molecules per spleen (10(8) cells). The same number of [125I]IgG molecules were bound to chromatin in cell cultures. The uptake of [125I]IgG was competitively inhibited by non-labelled IgG. Binding of [125I]IgG molecules reextracted from cytoplasm and chromatin with specific anti-human IgG serum argues against the uptake of degraded [125I]IgG molecules. [125I]IgG was tightly bound to DNA. Approximately 50 per cent of [125I]IgG was present in the residual chromatin fraction (after removal of 0.35 M and 2 M NaCl-soluble fractions) and 40 per cent was complexed with DNA (after removal of histones and non-histones AP1, AP2, AP3 and AP4). Binding of [125I]IgG by isolated chromatin was inhibited by the cytoplasmic fraction but not by BSA. Binding of [125I]IgG by fractionated chromatin, (chromatins remaining after removal of 0.35M, and 2M NaCl-soluble fractions or histones + non-histones AP1 + AP2 + AP3 + AP4) occurred at a level similar to that observed with native chromatin. The results suggest that interaction of antigen with immunocompetent cells is not restricted to the cell surface but that antigen seems to be taken up into cytoplasm, migrates to the nuclei and is bound to chromatin, probably directly to DNA. The results are discussed in relation to the induction of the immune reaction.

Incorporation of antigen ¹²⁵I IgG into particualr cell compartments: Binding by chromatin

Cell Biochemistry & Function

1985

Self Cite

Differences in salt solubility, DNase II sensitivity and template activity of chromatin from antibody synthesizing spleen cells and myeloma cells RPC 5 and ABPC 22

1982

Mol Cell Biochem

Chromatin from myeloma cells RPC5 and ABPC22, and from spleen and liver cells of immunized rats and mice, and mice bearing tumours, was fractionated into three part: 0.35 M NaCl-soluble, 2 M NaCl-soluble and residual. The residual fraction from myeloma cells differed from that of immunized spleen cells, described previously as containing unique sequences (5), in that it has higher protein and DNA levels, lower DNase II sensitivity and lower template activity.

Specific non-histone protein fractions associated with ‘active’ chromatin in immunized rats

1982

Mol Cell Biochem

Non-histone proteins of normal, non-immunized rats and rats immunized with mouse spleen cells were labelled with three different amino acids: [3H]tryptophan, [3H]methionine and [3H]leucine. Chromatin was fractionated at increasing salt concentrations into three fractions: 0.35 M NaCl-soluble, 2 M NaCl-soluble and residual. Non-histone protein fractions F (Mr 12 000) and H (Mr 3 000) highly labelled with [3H]tryptophan, lower with [3H]methionine but not with [3H]leucine, were present mainly in the residual fraction. After DNAse II treatment non-histone protein fractions F and H disappeared in chromatin fractions and were present in Mg2+ soluble fractions which suggests that, similar to the fractions I (Mr below 3 000) and B (Mr 120 000) described previously (5), these fractions may be associated with active transcribed genes.