Condensation of chromatin into chromosomes preserves an open configuration but alters the DNase I hypersensitive cleavage sites of the transcribed gene.
DNase I was used to probe the molecular organization of the chicken ovalbumin (OV) gene and glyceraldehyde 3-phosphate dehydrogenase (GPD) gene in interphase nuclei and in metaphase chromosomes of cultured chicken lymphoblastoid cells (MSB-1 line). The OV gene was not transcribed in this cell line, whereas the GPD gene was constitutively expressed. The GPD gene was more sensitive to DNase I digestion than the OV gene in both interphase nuclei and metaphase chromosomes, as determined by Southern blotting and liquid hybridization techniques. In addition, we observed DNase I hypersensitive sites around the 5' region of the GPD gene. These hypersensitive sites were not always at the same locations between the interphase nuclei and metaphase chromosomes. Our results suggest that chromatin condensation and decondensation during cell cycle alters nuclease hypersensitive cleavage sites.
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002645:Chickens; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure*; D002875:Chromosomes / Q000378:metabolism / Q000648:ultrastructure*; D003850:Deoxyribonuclease I; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005796:Genes; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D008214:Lymphocytes; D009693:Nucleic Acid Hybridization; D010047:Ovalbumin / Q000235:genetics*; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
D002843:Chromatin; D010047:Ovalbumin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I