DNAase hydrolysis of chromatin DNA in intact sea urchin sperm cells. Effect of the ionic strength on the digestion parameters. |
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| Authors: | G Geraci L Noviello |
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| Affiliation: | 1. Center for Reproductive Biology, School of Biological Sciences, Washington State University, Pullman, WA 99164-4236, USA;2. Department of Biology, University of Texas at San Antonio, San Antonio, TX 78249, USA;1. The Lundquist Institute for Biomedical Innovation at Harbor-UCLA, Torrance, CA 90502, USA;2. Department of Medical Genetics, China Medical University, Shenyang 110122, China;3. Department of Physiology and Cell Biology, University of Nevada, Reno School of Medicine, Reno, NV 89557, USA;4. Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA;1. Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058 Basel, Switzerland;2. Faculty of Sciences, University of Basel, 4056 Basel, Switzerland |
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| Abstract: | Chromatin DNA in intact functional Sphaerechinus granularis sperm cells has been digested with micrococcal nuclease at three different ionic strength conditions. The results show a highly-flexible chromatin organization similar to that found in sperm heads. The rate of digestion, the limit value of acid-soluble material and the fragmentation pattern show that the sensitivity of nucleosome and internucleosome DNA regions to nuclease hydrolysis depends on a delicate balance of polar and non polar interactions. At low ionic strength, both nucleosome and internucleosome regions are rapidly and completely hydrolysed at the same time and a transient subunit fragment of 120 b.p. average length is formed. At high ionic strength, internucleosome regions are preferentially hydrolysed; there is a limit digest value and a stable subunit fragment of 140 b.p. average length is formed. A supernucleosome organization in the high ionic strength environment of the sperm cells is suggested by the transient preferential formation of heptamers of nucleosome DNA fragments. |
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