DOI: 10.1101/gad.5.4.683. Transcription on nucleosomal templates by RNA polymerase II in vitro: inhibition of elongation with enhancement of sequence-specific pausing.

Transcription on nucleosomal templates by RNA polymerase II in vitro: inhibition of elongation with enhancement of sequence-specific pausing.

10.1101/gad.5.4.683

Crossref journal-article

Cold Spring Harbor Laboratory

Genes & Development (246)

Abstract

The process by which RNA polymerase II elongates RNA chains in vivo, where the template is at least partially in a nucleosomal configuration, remains poorly understood. To approach this question we have partially purified RNA polymerase II transcription complexes paused early in elongation. These complexes were then used as substrates for chromatin reconstitution. Elongation of the nascent RNA chains on these nucleosomal templates is severely inhibited relative to elongation on naked DNA templates. Elongation on the nucleosomal templates results in a reproducible template-specific pattern of transcripts generated by RNA polymerase pausing. The RNA polymerases are not terminated because the large majority will resume elongation upon the addition of Sarkosyl or 400 mM KCl. The effectiveness of RNA polymerase II pause/termination sites is enhanced by the presence of nucleosomes. For example, a pause site similar in sequence to the c-myc gene exon 1 terminator is used four to seven times more effectively in reconstituted templates. A comparison of elongation on templates bearing phased nucleosomes and on reconstituted templates that show no predominant phasing pattern indicates that the locations of pause sites are not related to the positions of the nucleosomes. Rather, the major determinant of RNA polymerase pausing on the nucleosomal templates appears to be the underlying DNA sequence.

Bibliography

Izban, M. G., & Luse, D. S. (1991). Transcription on nucleosomal templates by RNA polymerase II in vitro: inhibition of elongation with enhancement of sequence-specific pausing. Genes & Development, 5(4), 683â696.

Authors 2

M G Izban (first)
D S Luse (additional)

References 33 Referenced 200

10.1016/S0022-2836(05)80123-8
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10.1126/science.2158670
10.1016/0092-8674(90)90717-S
{'key': '2021111320171477000_5.4.683.8', 'first-page': '2390', 'article-title': 'Transcription factor requirements for in vitro formation of transcriptionally competent 5S rRNA gene chromatin.', 'volume': '10', 'year': '1990', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Transcription factor requirements for in vitro formation of transcriptionally competent 5S rRNA gene chromatin. (1990)
10.1016/0092-8674(84)90298-8
10.1016/S0021-9258(17)39577-7 / J. Biol. Chem. / Separation and partial characterization of three functional steps in transcription initiation by human RNA polymerase II. (1985)
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{'key': '2021111320171477000_5.4.683.13', 'first-page': '3114', 'article-title': 'Assembly of RNA polymerase II preinitiation complexes before assembly of nucleosomes allows efficient initiation of transcription on nucleosomal templates.', 'volume': '8', 'year': '1988', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Assembly of RNA polymerase II preinitiation complexes before assembly of nucleosomes allows efficient initiation of transcription on nucleosomal templates. (1988)
10.1016/0092-8674(87)90561-7
10.1016/0092-8674(87)90338-2
10.1016/S0021-9258(19)75925-0 / J. Biol. Chem. / Transcription initiation by RNA polymerase II in vitro. At least two nucleotides must be added to form a stable ternary complex. (1987)
10.1016/0076-6879(89)70051-3
{'key': '2021111320171477000_5.4.683.18', 'first-page': '1401', 'article-title': 'Transcription of adenovirus 2 major late and peptide IX genes under conditions of in vitro nucleosome assembly.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Transcription of adenovirus 2 major late and peptide IX genes under conditions of in vitro nucleosome assembly. (1987)
10.1002/j.1460-2075.1989.tb08362.x / EMBO J. / Nucleosomes inhibit both transcriptional initiation and elongation by RNA polymerase III in vitro. (1989)
10.1016/0092-8674(89)90399-1
10.1002/j.1460-2075.1990.tb08313.x / EMBO J. / Effect of transcription of yeast chromatin on DNA topology in vivo. (1990)
10.1016/S0021-9258(17)44836-8 / J. Biol. Chem. / In vitro evidence that transcription-induced stress causes nucleosome dissolution and regeneration. (1990)
10.1016/S0021-9258(18)61178-0 / J. Biol. Chem. / Purification and functional characterization of transcription factor SII from calf thymus. Role in RNA polymerase II elongation. (1987)
10.1016/S0021-9258(18)61507-8 / J. Biol. Chem. / Factors involved in specific transcription by mammalian RNA polymerase II. Transcription factor IIS stimulates elongation of RNA chains. (1987)
10.1016/S0021-9258(18)81692-1 / J. Biol. Chem. / Transcription elongation factor SII (TFIIS) enables RNA polymerase II to elongate through a block to transcription in a human gene in vitro. (1989)
10.1073/pnas.86.1.12
10.1016/S0021-9258(19)81605-8 / J. Biol. Chem. / Elongation by Drosophila RNA polymerase II. Transcription of 3′-extended DNA templates. (1988)
{'key': '2021111320171477000_5.4.683.28', 'first-page': '777', 'article-title': 'Transcription elongation and eukaryotic gene regulation.', 'volume': '5', 'year': '1990', 'journal-title': 'Oncogene'} / Oncogene / Transcription elongation and eukaryotic gene regulation. (1990)
10.1101/gad.4.1.75
10.1016/S0021-9258(17)42806-7 / J. Biol. Chem. / Early events in the stimulation of mammary tumor virus RNA synthesis by glucocorticoids. (1984)
10.1073/pnas.86.24.9817
10.1016/0092-8674(87)90130-9
10.1101/gad.3.11.1814

Dates

Type	When
Created	18 years, 2 months ago (June 5, 2007, 5:15 p.m.)
Deposited	3 years, 9 months ago (Nov. 13, 2021, 11:17 p.m.)
Indexed	1 month, 2 weeks ago (July 11, 2025, 6:50 a.m.)
Issued	34 years, 4 months ago (April 1, 1991)
Published	34 years, 4 months ago (April 1, 1991)
Published Online	34 years, 4 months ago (April 1, 1991)
Published Print	34 years, 4 months ago (April 1, 1991)

Funders 0

None

BibTeX

@article{Izban_1991, title={Transcription on nucleosomal templates by RNA polymerase II in vitro: inhibition of elongation with enhancement of sequence-specific pausing.}, volume={5}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.5.4.683}, DOI={10.1101/gad.5.4.683}, number={4}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Izban, M G and Luse, D S}, year={1991}, month=apr, pages={683–696} }

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