+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Synthetic sites for transcription termination and a functional comparison with tryptophan operon termination sites in vitro



Synthetic sites for transcription termination and a functional comparison with tryptophan operon termination sites in vitro



Proceedings of the National Academy of Sciences of the United States of America 78(7): 4180-4184



Termination of transcription by Escherichia coli RNA polymerase in vitro appears to depend primarily on 2 structural features of the termination site; a G + C(guanine, cytosine)-rich region of dyad symmetry and a series of terminal uridine residues in the transcript. To determine whether these 2 features are sufficient to specify .rho.-independent termination in vitro, new sequences were introduced within a tryptophan (trp) operon structural gene to create 2 sites with these characteristics. Transcription with wild-type RNA polymerase in vitro demonstrates that discrete termination occurs at one of these new sites, although at a low level. Use of the mutant RNA polymerase rpo203, which is more sensitive to certain weak terminators than is the wild-type enzyme, increases termination at both sites. The activity of our synthetic terminators was compared with those of several termination sites in the E. coli trp operon. Under normal conditions of transcription in vitro, termination becomes more efficient with an increase in the length of the stem in the RNA hairpin or an increase in the number of consecutive uridine residues. Transcription with the rpo203 polymerase and with ribonucleotide analogs gives changes consistent with these general trends. A model for termination involving separate but essential roles for the RNA hairpin and the stretch of uridines in the transcript is supported.

(PDF emailed within 0-6 h: $19.90)

Accession: 006573190

Download citation: RISBibTeXText

PMID: 7027254

DOI: 10.2307/10575


Related references

In vitro analysis of transcription initiation and termination from the Lhcb1 gene family in Nicotiana sylvestris: detection of transcription termination sites. Plant Journal 33(6): 1063-1072, 2003

Tandem termination sites in the tryptophan operon of Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 78(5): 2913-2917, 1981

Mutant rho factors with increased transcription termination activities. I. Functional correlations of the primary and secondary polynucleotide binding sites with the efficiency and site-selectivity of rho-dependent termination. Journal of Molecular Biology 210(1): 23-37, 1989

Transcription termination sites at the distal end of the leu operon of Salmonella typhimurium. Journal of Molecular Biology 194(3): 443-452, 1987

Transcription termination and processing sites in the bacteriophage lambda pL operon. Journal of Molecular Biology 189(1): 131-141, 1986

Chromatin structure of xenopus ribosomal dna transcription termination sites evidence for a 2 step process of transcription termination. Chromosoma 86(5): 703-715, 1982

Rho dependent termination of transcription 1. identification and characterization of termination sites for transcription from the bacterio phage lambda p r promoter. Journal of Biological Chemistry 258(15): 9553-9564, 1983

Chromatin structure of Xenopus rDNA transcription termination sites. Evidence for a two-step process of transcription termination. Chromosoma 86(5): 703-715, 1982

Termination of transcription in vitro in the Escherichia coli tryptophan operon leader region. Journal of Molecular Biology 103(2): 383-393, 1976

Transcription termination sites in the major leftward operon of coli phage lambda. Virology 88(2): 252-262, 1978

Tandem transcription-termination sites in the late rightward operon of bacteriophage lambda. Molecular and General Genetics 189(2): 289-297, 1983

Rho-dependent termination of transcription: identification and characterization of termination sites for transcription from the bacteriophage l PR promoter. The Journal of Biological Chemistry 258: 53-64, 1983

Rho factor tandem termination sites and rna processing are all required to generate the mature 3' end of escherichia coli tryptophan operon messenger rna. Journal of Cellular Biochemistry Supplement (9 PART B): 183, 1985

Transcription termination in vitro at the tryptophan operon attenuator is controlled by secondary structures in the leader transcript. Proceedings of the National Academy of Sciences of the United States of America 80(8): 2206-2210, 1983

The attenuator of the tryptophan operon in E.coli: rho-mediated release of RNA polymerase from a transcription termination complex in vitro. Nucleic Acids Research 5(12): 4613-4623, 1978