Abstract
Rat-liver nucleoli (10-15 pg DNA) were digested with either 0.6 or 3 units of DNase I for various times (up to 1 h). RNA synthesis was then measured in the absence or presence ol 3 units ofEscherichia coli RNA polymerase. It was found that the nucleolar chromatin supporting the endogenous engaged RNA polymerase I transcription was compl-etely destroyed in 3 min with either concentration of DNase I. The nucleolar chromatin template transcribed byE. coli RNA polymerase retained 50% of its original capacity even 60 min alter 3 units of DNase I digestion. When hybridization experiments were conducted, it was found that the DNAs derived from both levels of DNase-Idigested nucleoli were incapable of forming hybrids with the labelled nucleolar RNA synthesized by the engaged RNA polymerase I from the untreated nucleoli. Since the engaged RNA polymerase I transcribes only the physiologically active genes of the nucleolar chromatin, and the RNA transcripts represent active gene product, these data suggest that DNase I digestion has completely destroyed the active genes of the nucleolar chromatin, andE. coli RNA polymerase is able to transcribe the inactive nucleolar chromatin template.
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References
Keshgegian AA & Furth JJ (1972) Biochem. Biophys. Res. Commun.48, 757–763.
Keshgegian AA, Garbian GS & Furth JJ (1973) Biochemistry12, 4337–4342.
Ballal NR, Choi YC, Mouche R & Busch H (1977) Proc. Natl. Acad. Sci. U.S.A.74, 2446–2450.
Weintraub H & Groudine J (1976) Science193, 848–856.
Garel A & Axel R (1976) Proc. Natl. Acad. Sci. U.S.A.73, 3966–3970.
Mathis D, Dudet P & Chambon P (1980) Prog. Nucleic Acid Res.24, 1–55.
Yu FL (1981) J. Biol. Chem.256, 3292–3297.
Quagliarotti G, Hidregi E, Wikman V & Busch H (1970) J. Biol. Chem.245, 1962–1969.
Gillespie D & Spiegelman S (1965) J. Mol. Biol.12, 820–842.
Yu FL & Feigelson P (1971) Proc. Natl. Acad. Sci. U.S.A.68, 2177–2180.
Grummt I (1975) Eur. J. Biochem.57, 159–167.
Roeder RG (1976) inRNA Polymerase (Losick H & Chamberlin M, eds), pp. 285–329, Cold Spring Harbor Press, New York.
Yu FL (1974) Nature251, 344–346.
Yu FL (1975) Biochem. Biophys. Acta395, 329–336.
Zasloff M & Felsenfeld G (1977) Biochem. Biophys. Res. Commun.75, 598–603.
Zasloff M & Felsenfeld G (1977) Biochem.stry16, 5135–5145.
Giesecke K, Sippel AE, Nguyen-Hau NC, Groner B, Hynes NE, Wurtz T & Schultz G (1977) Nucleic Acids Res.4, 3943–3958.
Palmiter RD & Lee DC (1980) J. Biol. Chem.255, 9693–9698.
Reich E & Goldberg IH (1964) Prog. Nucleic Acid Res. of Molec. Biol.3, 183–234.
Yu FL (1980) Biochem. J.188, 381–385.
Allfrey VG (1980)Cell Biology—A Comprehensive Treatment (Goldstein L & Prescott DM, eds), vol. 3, pp 347–437, Academic Press, New York.
Razin A & Rings AD (1980) Science210, 604–610.
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Yu, FL., Barrett, A. Evidence for the transcription of physiologically inactive rat-liver nucleolar chromatin byEscherichia coli RNA polymerase. Biosci Rep 2, 155–161 (1982). https://doi.org/10.1007/BF01116378
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DOI: https://doi.org/10.1007/BF01116378