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On lysis of macronuclei from the ciliated protozoan Oxytricha at 0.5-2 M NaCl, the DNA, which is normally found as discrete molecules ranging from 0.5 to 20 kilobases, appears in high molecular weight aggregates. Various treatments of the macronuclear lysate (i.e., nucleases, proteases, variation of salt, pH, and temperature) indicate that preservation of the aggregate structure depends on both nucleic acid-nucleic acid and nucleic acid-protein interactions. Purification of the DNA-protein complex after lysing the nuclei in 2 M NaCl shows that one major nuclear protein copurifies with the DNA. As shown by DNA-protein binding experiments, this protein has a high affinity for DNA; however, no evidence for sequence specificity of the protein binding was obtained. Chromatin reconstitution experiments suggest that the protein in itself is not sufficient for DNA aggregation in nuclei, but other factors, possibly the native chromatin structure, are required. Electron microscopy of the purified DNA-protein complex showed structures similar to those observed previously with in vitro-aggregated purified macronuclear DNA (14). A model is presented in which the terminal inverted repeat sequences found on all macronuclear DNA molecules interact with each other forming multistranded DNA complexes. The formation of these structures may be accelerated and stabilized by a protein in vivo.

Macronuclear DNA of hypotrichous ciliates exists in the form of gene-sized DNA molecules. It can be resolved on agarose gels into a continuum of sizes upon which is imposed a set of characteristic DNA bands. Most or all of the DNA molecules carry identical terminal inverted repeat sequences. By incubating macronuclear DNA under increasingly stronger ionic conditions, high molecular weight DNA aggregates and ring-like DNA structures are formed. Experimental evidence is presented that this aggregation is not due to the presence of identical single-stranded DNA ends on each macronuclear DNA fragment, and an alternative model for DNA aggregation is discussed.