Diffusible factors are responsible for differences in nuclease sensitivity among chromatins originating from different cell types.
We have examined the kinetics of nuclease digestion of chromatin from committed and uncommitted cells in experiments where the nuclei are mixed and co-digested. Cultures of the sea urchin, Arbacia punctulata, were grown to the 16-cell stage in either [3H]thymidine or [14C]thymidine and the macromere, mesomere, and micromere cell types separated. After isolation, sets of nuclei with two different blastomere types (each having different radionucleotide tagging) were mixed and co-digested with micrococcal nuclease or DNase. I. The extent of digestion was monitored by solubility in 5% perchloric acid (PCA). We find no significant differences in initial digestion rates or limit digests among the different cell types when co-digested with either nuclease. Differences in nuclease sensitivity observed when nuclei are digested separately are abolished when nuclei are probed in a mixing experiment. The results support the hypothesis that phenotypic differences in digestibility among different cell types in vitro reflect differences in chromatin-condensing factors which can diffuse between nuclei.
D000818:Animals; D001757:Blastomeres / Q000166:cytology; D002250:Carbon Radioisotopes; D002455:Cell Division; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000502:physiology / Q000648:ultrastructure*; D004261:DNA Replication; D003850:Deoxyribonuclease I; D004625:Embryo, Nonmammalian / Q000166:cytology; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005260:Female; D007700:Kinetics; D008836:Micrococcal Nuclease / Q000378:metabolism*; D012617:Sea Urchins / Q000196:embryology; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D002250:Carbon Radioisotopes; D002843:Chromatin; D014316:Tritium; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease; D013936:Thymidine