Abstract
Nuclei of mouse liver and an immunoglobulin producing myeloma were digested with HaeIII or its isoschizomer BspRI. The DNA fragment patterns after electrophoresis, blotting, and hybridization were very similar for the two types of nuclei when a probe for the non-expressed beta-globin gene was used. The constant (C) region of the kappa light chain gene, on the other hand, was more accessible to the nuclease in myeloma than on liver nuclei. In myeloma nuclei the BspRI sites were about equally sensitive, in fact the pattern resembled a partial digestion pattern of free DNA. In contrast, in liver nuclei some sites were much more protected than others. This is interpreted by assuming that this single copy non-expressed gene region is covered by nucleosomes which are preferentially located on certain DNA sequences. Restriction nuclease digestion of nuclei seems to be a promising method for the analysis of genes in their expressed and non-expressed states.
Full text
PDF

















Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bellard M., Kuo M. T., Dretzen G., Chambon P. Differential nuclease sensitivity of the ovalbumin and beta-globin chromatin regions in erythrocytes and oviduct cells of laying hen. Nucleic Acids Res. 1980 Jun 25;8(12):2737–2750. doi: 10.1093/nar/8.12.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bloom K. S., Anderson J. N. Fractionation of hen oviduct chromatin into transcriptionally active and inactive regions after selective micrococcal nuclease digestion. Cell. 1978 Sep;15(1):141–150. doi: 10.1016/0092-8674(78)90090-9. [DOI] [PubMed] [Google Scholar]
- Curtis P. J., Mantei N., Weissmann C. Characterization and kinetics of synthesis of 15S beta-globin RNA, a putative precursor of beta-globin mRNA. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):971–984. doi: 10.1101/sqb.1978.042.01.098. [DOI] [PubMed] [Google Scholar]
- Gottesfeld J. M., Garrard W. T., Bagi G., Wilson R. F., Bonner J. Partial purification of the template-active fraction of chromatin: a preliminary report. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2193–2197. doi: 10.1073/pnas.71.6.2193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gould D. C., Callan H. G., Thomas C. A., Jr The actions of restriction endonucleases on lampbrush chromosomes. J Cell Sci. 1976 Jul;21(2):303–313. doi: 10.1242/jcs.21.2.303. [DOI] [PubMed] [Google Scholar]
- Greene P. J., Heyneker H. L., Bolivar F., Rodriguez R. L., Betlach M. C., Covarrubias A. A., Backman K., Russel D. J., Tait R., Boyer H. W. A general method for the purification of restriction enzymes. Nucleic Acids Res. 1978 Jul;5(7):2373–2380. doi: 10.1093/nar/5.7.2373. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hamlyn P. H., Browniee G. G., Cheng C. C., Gait M. J., Milstein C. Complete sequence of constant and 3' noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencing. Cell. 1978 Nov;15(3):1067–1075. doi: 10.1016/0092-8674(78)90290-8. [DOI] [PubMed] [Google Scholar]
- Hewish D. R., Burgoyne L. A. Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease. Biochem Biophys Res Commun. 1973 May 15;52(2):504–510. doi: 10.1016/0006-291x(73)90740-7. [DOI] [PubMed] [Google Scholar]
- Hörz W., Igo-Kemenes T., Pfeiffer W., Zachau H. G. Specific cleavage of chromatin by restriction nucleases. Nucleic Acids Res. 1976 Nov;3(11):3213–3226. doi: 10.1093/nar/3.11.3213. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Igo-Kemenes T., Greil W., Zachau H. G. Prepartation of soluble chromatin and specific chromatin fractions with restriction nucleases. Nucleic Acids Res. 1977 Oct;4(10):3387–3400. doi: 10.1093/nar/4.10.3387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Igó-Kemenes T., Zachau H. G. Domains in chromatin structure. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):109–118. doi: 10.1101/sqb.1978.042.01.012. [DOI] [PubMed] [Google Scholar]
- Kiss A., Sain B., Csordás-Tòth E., Venetianer P. A new sequence-specific endonuclease (Bsp) from Bacillus sphaericus. Gene. 1977 Jul;1(5-6):323–329. doi: 10.1016/0378-1119(77)90037-3. [DOI] [PubMed] [Google Scholar]
- Kornberg R. D. Structure of chromatin. Annu Rev Biochem. 1977;46:931–954. doi: 10.1146/annurev.bi.46.070177.004435. [DOI] [PubMed] [Google Scholar]
- Lipchitz L., Axel R. Restriction endonuclease cleavage of satellite DNA in intact bovine nuclei. Cell. 1976 Oct;9(2):355–364. doi: 10.1016/0092-8674(76)90125-2. [DOI] [PubMed] [Google Scholar]
- Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perry R. P., Kelley D. E., Coleclough C., Seidman J. G., Leder P., Tonegawa S., Matthyssens G., Weigert M. Transcription of mouse kappa chain genes: implications for allelic exclusion. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1937–1941. doi: 10.1073/pnas.77.4.1937. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pfeiffer W., Horz W., Igo-Kemenes T., Zachau H. G. Restriction nucleases as probes of chromatin structure. Nature. 1975 Dec 4;258(5534):450–452. doi: 10.1038/258450a0. [DOI] [PubMed] [Google Scholar]
- Prunell A., Kornberg R. D. Relation of nucleosomes to DNA sequences. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):103–108. doi: 10.1101/sqb.1978.042.01.011. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Stalder J., Groudine M., Dodgson J. B., Engel J. D., Weintraub H. Hb switching in chickens. Cell. 1980 Apr;19(4):973–980. doi: 10.1016/0092-8674(80)90088-4. [DOI] [PubMed] [Google Scholar]
- Steinmetz M., Zachau H. G., Mach B. Cloning of immunoglobulin kappa light chain genes from mouse liver and myeloma MOPC 173. Nucleic Acids Res. 1979 Jul 25;6(10):3213–3229. doi: 10.1093/nar/6.10.3213. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinmetz M., Zachau H. G. Two rearranged immunoglobulin kappa light chain genes in one mouse myeloma. Nucleic Acids Res. 1980 Apr 25;8(8):1693–1707. doi: 10.1093/nar/8.8.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Varshavsky A. J., Sundin O., Bohn M. A stretch of "late" SV40 viral DNA about 400 bp long which includes the origin of replication is specifically exposed in SV40 minichromosomes. Cell. 1979 Feb;16(2):453–466. doi: 10.1016/0092-8674(79)90021-7. [DOI] [PubMed] [Google Scholar]
- Weintraub H., Groudine M. Chromosomal subunits in active genes have an altered conformation. Science. 1976 Sep 3;193(4256):848–856. doi: 10.1126/science.948749. [DOI] [PubMed] [Google Scholar]
- Wu C., Bingham P. M., Livak K. J., Holmgren R., Elgin S. C. The chromatin structure of specific genes: I. Evidence for higher order domains of defined DNA sequence. Cell. 1979 Apr;16(4):797–806. doi: 10.1016/0092-8674(79)90095-3. [DOI] [PubMed] [Google Scholar]
- Wu C., Wong Y. C., Elgin S. C. The chromatin structure of specific genes: II. Disruption of chromatin structure during gene activity. Cell. 1979 Apr;16(4):807–814. doi: 10.1016/0092-8674(79)90096-5. [DOI] [PubMed] [Google Scholar]