ADP-Ribosylated Histone H1 from HeLa Cultures. Fundamental Differences to (ADP-Ribose)n-Histone H1 Conjugates Formed in vitro

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Abstract
ADP-ribosylated histone H1 was isolated from intact HeLa (human cervical cancer) cells grown for 24 h with [3H]-adenosine and compared with ADP-ribosylated histone H1 synthesized from [3H]NAD by isolated HeLa nuclei. Most (ADP-ribose)n-histone H1 conjugates formed in vivo carried single ADP-ribose units, less than 1/4 of the total ADP-ribose residues being in the form of oligomeric or polymeric chains. (ADP-ribose)n linked to H1 in vivo was not released by neutral NH2OH to a significant extent. Alkali treatment (pH 10.5) liberated most but not all of the ADP-ribose residues which may indicate the existence of a new type of linkage so far found only in conjugates isolated from intact tissue. No ADP-ribosylated histone H1 complex of higher MW (H1 dimer) was detected in intact cells. By contrast, isolated HeLa nuclei formed ADP-ribosylated histone H1 which contained predominantly polymeric ADP-ribose residues. The (ADP-ribose)n residues were linked by NH2OH-sensitive and by NH2OH-resistant, alkali (pH 10.5) labile bonds, the majority of the conjugates appearing in the form of the higher-MW complex. A comparison with the ADP-ribosylated non-histone proteins indicated that histone H1 formed in vivo carried less than 2.5% of the total protein-bound ADP-ribose residues and less than 1% of the protein-bound ADP-ribose synthesized in vitro.