The association of human c-Ha-ras sequences with chromatin and nuclear proteins.
As a step towards the understanding of possible relationship between chromatin organization and regulation of the oncogene expression, we have investigated the chromatin structure of one of the more frequently activated oncogenes, c-Ha-ras, in HeLa-S3 cells. This was accomplished by isolation of the chromatin fractions (soluble and insoluble) after micrococcal nuclease digestion of purified nuclei and probing for the distribution of ras sequences. The polynucleosomal fraction was further resolved by sucrose gradient sedimentation. Southern-blot hybridization of the DNA isolated from various fractions yielded following results: (1) c-Ha-ras sequences segregated predominantly in the lysate fraction. (2) Unlike the B-globin (transcriptionally inactive) sequences, ras-H associated chromatin lacked typical nucleosomal packaging. Furthermore, since post-translational modifications of nuclear proteins have been suggested to modulate the nucleosome structure during DNA transcription and replication, ras sequences, in polynucleosomes immunofractionated on anti-poly (ADP-Ribose) Sepharose were also examined. The data suggested that the major class of this oncogene sequence exists in chromatin more distal to the sites of this particular chromatin modification.
D001483:Base Sequence; D002843:Chromatin / Q000032:analysis*; D005786:Gene Expression Regulation; D006367:HeLa Cells / Q000032:analysis; D006801:Humans; D007163:Immunosorbent Techniques; D008836:Micrococcal Nuclease / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D009698:Nucleoproteins / Q000032:analysis*; D009857:Oncogenes; D011064:Poly Adenosine Diphosphate Ribose / Q000276:immunology; D014158:Transcription, Genetic
D002843:Chromatin; D009698:Nucleoproteins; D011064:Poly Adenosine Diphosphate Ribose; D008836:Micrococcal Nuclease