title: Globin RNA synthesis in vitro by isolated erythroleukemic cell nuclei: direct evidence for increased transcription during erythroid differentiation.
abstract: Murine erythroleukemic cells accumulate cytoplasmic globin mRNA during differentiation induced in tissue culture by dimethyl sulfoxide. Cellular accumulation of globin RNA may reflect transcriptional activation of the globin genes and/or posttranscriptional stabilization of globin RNA during differentiation. To evaluate possible transcriptional controls directly; globin RNA synthesis by isolated erythroleukemic cell nuclei was studied. Conditions were established for optimal nuclear RNA synthesis in vitro in the presence of a mercurinucleotide (Hg-CTP). Mercurated RNA synthesized in vitro was purified free of endogenous RNA by affinity chromatography on sulfhydryl-Sepharose, and analyzed for the presence of newly synthesized globin RNA sequences by molecular hybridization to globin complementary [32P]DNA. The results demonstrate markedly increased synthesis of globin RNA by nuclei isolated from dimethyl sulfoxide-treated cells, even within 5 min of nuclear transcription in vitro. These findings are most consistent with transcriptional activation of the globin genes upon induction of differentiation.
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005914:Globins / Q000096:biosynthesis*; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000096:biosynthesis*; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D012333:RNA, Messenger; D013438:Sulfhydryl Compounds; D005914:Globins; D004121:Dimethyl Sulfoxide
references.citation: J Mol Biol. 1976 Oct 5;106(4):1067-75
references.pmid: 824454
references.citation: J Biol Chem. 1970 Jun;245(12):3059-65
references.pmid: 5432796
references.citation: Proc Natl Acad Sci U S A. 1972 Dec;69(12):3620-3
references.pmid: 4509323
references.citation: Biochemistry. 1974 Apr 23;13(9):1896-9
references.pmid: 4857734
references.citation: J Mol Biol. 1973 Oct 25;80(2):229-41
references.pmid: 4587403
references.citation: Proc Natl Acad Sci U S A. 1973 Aug;70(8):2238-42
references.pmid: 4365367
references.citation: Nat New Biol. 1971 Sep 29;233(39):134-6
references.pmid: 5286748
references.citation: Cell Differ. 1974 Jun;3(1):9-22
references.pmid: 4136752
references.citation: Proc Natl Acad Sci U S A. 1971 Nov;68(11):2861-5
references.pmid: 4108111
references.citation: Biochemistry. 1975 Jun 3;14(11):2447-57
references.pmid: 1138868
references.citation: Proc Natl Acad Sci U S A. 1975 Jan;72(1):98-102
references.pmid: 164031
references.citation: Proc Natl Acad Sci U S A. 1975 Dec;72(12):4835-9
references.pmid: 1108006
references.citation: J Biol Chem. 1975 Nov 25;250(22):8753-60
references.pmid: 1237491
references.citation: Cell. 1976 Aug;8(4):495-503
references.pmid: 954102
references.citation: Cell. 1976 Feb;7(2):267-77
references.pmid: 986246
references.citation: Proc Natl Acad Sci U S A. 1976 May;73(5):1564-7
references.pmid: 1064024
references.citation: N Engl J Med. 1976 Sep 23;295(13):710-4
references.pmid: 785253
references.citation: Proc Natl Acad Sci U S A. 1976 Jun;73(6):1811-5
references.pmid: 1064854
references.citation: Proc Natl Acad Sci U S A. 1976 Mar;73(3):775-9
references.pmid: 815908
references.citation: Biochemistry. 1976 Jul 27;15(15):3270
references.pmid: 986156
references.citation: J Mol Biol. 1972 Jun 28;67(3):433-41
references.pmid: 4558099
references.citation: Biochemistry. 1975 Jun 3;14(11):2458-69
references.pmid: 166657
references.citation: Cell. 1974 Jul;2(3):197-204
references.pmid: 4415110
references.citation: Biochemistry. 1973 Aug 28;12(18):3440-6
references.pmid: 4731189
references.citation: Proc Natl Acad Sci U S A. 1971 Feb;68(2):378-82
references.pmid: 5277089
references.citation: Proc Natl Acad Sci U S A. 1975 Aug;72(8):2930-4
references.pmid: 1059083
references.citation: Biochemistry. 1976 Feb 24;15(4):824-9
references.pmid: 1247536
references.citation: Dev Biol. 1976 Jan;48(1):118-31
references.pmid: 1245255
references.citation: Biochem Biophys Res Commun. 1977 Apr 11;75(3):598-603
references.pmid: 324468