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Nucleic Acids Research, 1995, Vol. 23, No. 6 1075-1082
© 1995

MOLECULAR BIOLOGY

Deposition of histone H1 onto reconstituted nucleosome arrays inhibits both initiation and elongation of transcripts by 17 RNA polymerase

Timothy E. O'Neill1,*, Geert Meersseman1,+, Sari Pennings1,+ and E. Morton Bradbury1,2

1Department of Biological Chemistry, School of Medicine, University of California Davis, CA 95616, USA 2Life Sciences Division, Los Alamos National Laboratory Los Alamos, NM 87545, USA

*To whom correspondence should be addressed

Received June 13, 1994. Revised December 16, 1994. Accepted December 16, 1994.

The effect of histone H1 on transcription by bacterio-phage T7 RNA polymerase was examined using reconstituted chromatin templates. A 3.8 kb linear DNA template consisting of a specific transcription promoter for T7 RNA polymerase placed upstream of 18 tandem repeats of a 207 bp nucleosome positioning sequence derived from the 5S rRNA gene of Lytechinusvarlegatus was used as a template for chromatin reconstitution. Regularly spaced arrays of nucleosome cores were assembled onto this DNA template from donor histone octamers by salt step dialysis. Histone H1 was incorporated onto free DNA or reconstituted chromatin templates and double label transcription assays were performed. The experiments indicated that histone H1 has a strong inhibitory effect on both transcription initiation and elongation. These effects are especially pronounced on chromatin templates, where both transcription initiation and elongation are virtually halted. The inhibition of transcription elongation appears to result from a dramatic increase in premature termination of transcripts. These experiments indicate that assembly of histone H1 into chromatin can result in structures which are completely repressed with respect to transcription.

+Present address: Department of Biochemistry, University of Edinburgh, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK


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