Novobiocin inhibits DNA topoisomerases. It also inhibits excision repair of DNA photodamage, blocking both repair synthesis and the earlier step of incision at u.v. damage sites (as measured by the accumulation of DNA strand breaks in u.v.-irradiated interphase cells treated with DNA synthesis inhibitors such as hydroxyurea or cytosine arabinoside). It has been supposed, therefore, that novobiocin affects repair by blocking a putative topoisomerase step prior to incision. But we find that novobiocin also has a marked dose- and time-dependent effect on mitochondria: in cells exposed to novobiocin, mitochondria swell and their cristae become disrupted, and the intracellular ATP:ADP ratio is lowered, though the membrane potential is maintained as judged by rhodamine 123 fluorescence. Mitotic cells are more resistant to mitochondrial disruption by novobiocin than are interphase cells. This correlates with a relative resistance of u.v.-irradiated mitotic cells to the inhibition of incision by novobiocin. The chromosomal decondensation that results from the accumulation of DNA breaks due to incision when u.v.-irradiated mitotic cells are treated with hydroxyurea and cytosine arabinoside is largely suppressed by novobiocin. Furthermore, the suppression of induced strand break accumulation is partly due to a suppression by novobiocin of the uptake and phosphorylation of cytosine arabinoside; breaks accumulated in u.v.-irradiated cells in the presence of aphidicolin, an inhibitor of DNA polymerase alpha that does not require phosphorylation, are less novobiocin-sensitive. We conclude that the effects of novobiocin on excision repair are more likely to be due to a non-specific effect on ATP metabolism than to a specific effect on a repair-related topoisomerase.

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A detailed electron microscopy study of cadmium sensitive and resistant muntjac fibroblast cell lines has identified a wide range of intracellular damage following exposure to cadmium. Damaged organelles included cell membrane, mitochondria, Golgi cisternae and tubular network, chromatin, nucleoli, microfilaments and ribosomes. Although cell membrane damage was generally the earliest indication of adverse cadmium action, particularly with continuous cadmium exposures, cells could tolerate extensive membrane loss. Mitochondrial distortion and some damage to Golgi was also tolerated. The turning point at which cadmium became lethal was generally marked by a cascade of events which included damage to both nuclear and cytoplasmic components. These results for fibroblasts are discussed and compared with damage reported in other types of cells.

Microinjection of adenine nucleotides and substrates into the cytoplasm of Amoeba proteus followed by EM examination has been used in an attempt to relate alterations in mitochondrial morphology with functional changes. Contracted mitochondria with dark matrix and wide cristae (Type I), and expanded mitochondria with light matrix and narrower cristae (Type II) coexist in normal active amoebae, but their numbers can be varied according to different cell activity states. Following injection of ATP, the mitochondria of the amoebae showed a time-dependent movement towards a predominately Type II form, whilst injections of ADP produced predominately the Type I form. Injection of succinate or deionised water, even in large amounts, had little effect on the numbers of Type I and Type II forms. The change induced by ATP was of long duration; that induced by ADP was influenced by both concentration injected and the cell's substrate levels. With 3 mM solutions of ADP the mitochondrial population was primarily of Type I organelles; higher ADP concentrations or the simultaneous injection of succinate, however, resulted in a switch with time to increased proportions of Type II mitochondria. The results extend the findings of previous in vivo and in vitro mitochondrial studies and are discussed in the light of these.