Two exocellular nucleases with molecular masses of 18 and 34 kDa, which are nutritionally regulated and reach their maximum activity during aerial mycelium formation and sporulation, have been detected in Streptomyces antibioticus. Their function appears to be DNA degradation in the substrate mycelium, and in agreement with this proposed role the two nucleases cooperate efficiently with a periplasmic nuclease previously described in Streptomyces antibioticus to completely hydrolyze DNA. The nucleases cut DNA nonspecifically, leaving 5-phosphate mononucleotides as the predominant products. Both proteins require Mg 2؉ , and the additional presence of Ca 2؉ notably stimulates their activities. The two nucleases are inhibited by Zn 2؉ and aurin tricarboxylic acid. The 18-kDa nuclease from Streptomyces is reminiscent of NUC-18, a thymocyte nuclease proposed to have a key role in glucocorticoid-stimulated apoptosis. The 18-kDa nuclease was shown, by aminoterminal protein sequencing, to be a member of the cyclophilin family and also to possess peptidylprolyl cistrans-isomerase activity. NUC-18 has also been shown to be a cyclophilin, and "native" cyclophilins are capable of DNA degradation. The S. antibioticus 18-kDa nuclease is produced by a proteolytic processing from a less active protein precursor. The protease responsible has been identified as a serine protease that is inhibited by N ␣ -ptosyl-L-lysine chloromethyl ketone and leupeptin. Inhibition of both of the nucleases or the protease impairs aerial mycelium development in S. antibioticus. The biochemical features of cellular DNA degradation during Streptomyces development show significant analogies with the late steps of apoptosis of eukaryotic cells.The actinomycetes are a large group of filamentous bacteria that are adapted for growth in soil by forming a ramifying network, called a mycelium. Within this group the predominant isolates belong to the genus Streptomyces, which produce a well developed branched mycelium on agar plates, resulting in a compact colony. In the vegetative phase, the filaments often lack cross-walls (substrate mycelium) and thus have several copies of the chromosome. When the colony ages, a characteristic aerial mycelium is formed, in response to unknown signals involving nutrient limitation, which subsequently fragment and/or sporulate by the synchronous formation of crosswalls in the multinucleate sporophores followed by separation of the individual cells directly into spores. This is similar to the growth and differentiation of fungi, and from the morphological and metabolic points of view, Streptomyces can be considered as boundary organisms (1). Coincident with the morphological differentiation, the streptomycetes produce numerous compounds (secondary metabolites) within which antibiotics are of commercial relevance. As corresponds to their habitat, these bacteria are nutritionally quite versatile, and most produce extracellular hydrolytic enzymes that permit the utilization of polysaccharides, proteins, fats, and other substrat...