Discrimination of several classes of nonhistone proteins in chromatin fractions from liver and thymus nuclei of rats.
Nonhistone proteins (NHPs) of salt-soluble chromatin (Chromatin S) and of the residual nuclei (Chromatin P) from rat liver and thymus were studied by SDS-polyacrylamide gel electrophoresis. The two chromatin fractions of the liver showed significant differences in their NHP patterns with most of the hnRNP and matrix proteins occurring in Chromatin P. In accordance with the low protein content of thymus nuclei, the corresponding thymus fractions exhibited electrophoretic patterns with a markedly lower amount of NHPs than in liver. Chromatin P from thymus, in contrast to the liver fraction, revealed only a very low content of hnRNP-specific proteins of molecular weight 30,000-40,000 (30 K to 40 K) (informosomal proteins) consistent with the significantly lower RNA content of thymus nuclei. In the region of the matrix proteins (60-75 K) Chromatin P showed only two bands of about 64 K and 73 K in thymus, whereas in liver five strong bands at 64 K, 66 K, 69 K, 73 K, and 75 K were found. RNase digestion was employed to discriminate hnRNP-specific protein from "real" chromosomal NHPs. At least about 65% and 25% of the NHPs from Chromatin P and S of liver, respectively, were found to be RNP-specific. The two chromatin fractions were further fractionated by sucrose gradient centrifugation and isopycnic banding in metrizamide. After centrifugation the main peaks, both of Chromatin S and P, contained only minor amounts of NHPs with a predominating protein of 38 K. By the centrifugation procedures described in this paper, a small subfraction of chromatin could be separated which was enriched in newly synthesized RNA, informosomal proteins, matrix- and other high molecular weight proteins. This subfraction might be related to transcriptionally active chromatin.
D000818:Animals; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000032:analysis*; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000032:analysis*; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000032:analysis*; D008854:Microscopy, Electron; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D012261:Ribonucleoproteins / Q000032:analysis; D013950:Thymus Gland / Q000032:analysis*
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D012261:Ribonucleoproteins