In vivo analysis of chromatin following nystatin-mediated import of active enzymes into Saccharomyces cerevisiae.
In vivo DNA-protein interactions are usually studied at the molecular level using DNA-degrading agents of low molecular weight. In order to be useful, macromolecular probes of chromatin structure, such as enzymes must first cross the cell membrane. In this paper we describe the introduction and evaluation of macromolecules with enzymatic activity into yeast spheroplasts treated with the polyene antibiotic nystatin. We report the low resolution analysis of chromatin structure in the promoter region of the Saccharomyces cerevisiae gene encoding DNA topoisomerase I by this technique using micrococcal nuclease and restriction enzymes.
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Fondazione Istituto Pasteur-Fondazione Cenci-Bolognetti, Roma, Italy.
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D001483:Base Sequence; D002462:Cell Membrane / Q000187:drug effects; D002463:Cell Membrane Permeability; D002843:Chromatin / Q000737:chemistry*; D004262:DNA Restriction Enzymes; D004264:DNA Topoisomerases, Type I / Q000235:genetics; D004271:DNA, Fungal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D005800:Genes, Fungal / Q000235:genetics*; D008836:Micrococcal Nuclease; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D009761:Nystatin / Q000494:pharmacology*; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013104:Spheroplasts / Q000187:drug effects
D002843:Chromatin; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D015345:Oligonucleotide Probes; D009761:Nystatin; D004262:DNA Restriction Enzymes; D008836:Micrococcal Nuclease; D004264:DNA Topoisomerases, Type I