RNA associated with non-histone chromosomal proteins in rat liver and in ascites hepatoma.
The purpose of the experiments was to determine if changes in the post-transcriptional processing of RNA in the hepatoma also affect the low molecular weight nuclear RNA which is transcribed from families of related genes and associated with non-histone chromosomal proteins. Separation of non-histone chromosomal proteins on Sephadex G-200 into three fractions separated the low molecular weight RNA associated with these proteins into metabolically stable RNA firmly bound to the first eluted fractions of high molecular weight non-histone chromosomal proteins, and into a metabolically active RNA which is eluted with the third protein fraction and can be separated from the low molecular weight non-histone chromosomal proteins by chromatography on DEAE-Sephadex A 25 (fraction III RNA). In liver as well as in hepatoma, this fraction III RNA represents about 50% of the RNA associated with the non-histone chromosomal proteins. Fraction III RNA from both tissues has an approximate molecular weight of 13,000, is rich in guanylic acid, is lacking dihydropyrimidines and is copied from the repetitive sequences of DNA. The content of uridylic acid is much higher in fraction III RNA isolated from hepatoma than in the same RNA isolated from liver, and competitive hybridization has shown that hepatoma fraction III RNA contains not only new base sequences which are not present in liver fraction III RNA, but also lacks some sequences which are present in the liver RNA. The technique of RNA/DNA hybridization in the presence of competing RNA has shown that, in hepatoma, the cytoplasmic RNA competes with more than 60% of the fraction III RNA for the hybridization sites on repetitive DNA. No competition was found when liver cytoplasmic RNA was used. The low ratio of competing hepatoma cytoplasmic RNA or of liver or hepatoma nuclear RNA which is required to displace fraction III RNA from its hybridization with DNA indicates that this RNA is synthesized and in hepatoma is also released into the cytoplasm as a part of larger RNA molecules. The detection of the nucleotide sequences found in liver associated with non-histone chromosomal proteins in the cytoplasm of hepatoma cells is evidence for extensive disruption of the post-transcriptional control in hepatoma.
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D005591:Chemical Fractionation; D002875:Chromosomes / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D005260:Female; D008113:Liver Neoplasms / Q000378:metabolism*; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D009698:Nucleoproteins / Q000032:analysis / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D014158:Transcription, Genetic
D009698:Nucleoproteins; D012334:RNA, Neoplasm