Excision Close to Matrix Attachment Regions of the Entire Chicken α- Globin Gene Domain by Nuclease S1 and Characterization of the Framing Structures

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 91; no. 10; pp. 4422 - 4426
• Main Authors: Targa, Felix Recillas, Razin, Sergey V., Claudia V. De Moura Gallo, Scherrer, Klaus
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 10-05-1994; National Acad Sciences
• Subjects: Animals; Base Sequence; Binding Sites; Cell Nucleus - metabolism; Cells, Cultured; Chickens - genetics; Deoxyribonuclease I; DNA; DNA - genetics; DNA - metabolism; DNA Primers; DNA probes; DNA-Binding Proteins - metabolism; Enzymes; Erythroblasts - metabolism; Erythroid cells; Genes; Genome; Genomes; Genomics; Globins - biosynthesis; Globins - genetics; Molecular Sequence Data; Nuclear matrix; Nucleotide sequences; Replication origin; Restriction Mapping; Single-Strand Specific DNA and RNA Endonucleases
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.10.4422

Nuclease S1-hypersensitive sites in a 40-kb region of the chicken genome including the domain of the α-globin genes were mapped. Brief treatment of isolated chicken erythroid cell nuclei with nuclease S1 allowed separation of an ≈20-kb genomic DNA fragment containing the whole α-globin gene cluster. No S1-hypersensitive sites were observed in the internal/part of the domain. The upstream S1 site was found in a DNA fragment of 1.7 kb where the origin of replication and several protein binding sites were identified previously. Precise mapping of the positions of S1 cleavage in this fragment and "in vivo" footprinting of DNA-protein interactions in isolated nuclei showed a correspondence with some of these protein binding sites. The possible significance of all these observations is discussed in connection with the replication origin and the nuclear matrix attachment regions in the framing structures.