Regulation of intrinsic terminator by translation in Escherichia coli

Transcription termination at a distance downstream

Hiroyuki Abe, Tatsuhiko Abo, Hiroji Aiba

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Background: Rho-independent terminators in Escherichia coli are DNA sequences of 30-50 bp consisting of a GC-rich dyad symmetry sequence followed by a run of T residues in the nontemplate strand. The transcription termination at the Rho-independent terminator occurs within the T-tract in vitro. It has been believed that the transcription termination at the Rho- independent terminator occurs within the T-tract in vivo, as established in vitro, and therefore the 3' ends of mRNAs are mostly generated as a direct result of transcription termination. However, how the transcription termination occurs and how the 3' ends of mRNAs are formed in living cells remains to be studied. Results: We developed a double terminator system in which a second Rho-independent terminator was placed downstream of the crp terminator. This system made it possible to detect transcripts that pass through the crp terminator by Northern blotting. We found that most of the crp transcripts extend beyond the crp terminator. The transcriptional read- through at the crp terminator was reduced when the translation of crp mRNA was interrupted. The level of the read-through transcript decreased with distance between the two terminators, suggesting that transcription termination occurs at multiple positions beyond the crp terminator. Conclusion: We conclude that most RNA polymerase reads through the crp terminator in the natural situation and terminates transcription over a wide region downstream of the crp terminator, resulting in heterogeneous primary transcripts that are subsequently processed back to the terminator hairpin. We propose that ribosome translation to the crp stop codon causes read- through of the terminator. The regulatory effect of translation on Rho- independent termination may be a general phenomenon at other operons.

Original languageEnglish
Pages (from-to)87-97
Number of pages11
JournalGenes to Cells
Volume4
Issue number2
Publication statusPublished - 1999
Externally publishedYes

Fingerprint

Escherichia coli
Messenger RNA
Terminator Codon
Protein Biosynthesis
DNA-Directed RNA Polymerases
Operon
Ribosomes
Northern Blotting
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Regulation of intrinsic terminator by translation in Escherichia coli : Transcription termination at a distance downstream. / Abe, Hiroyuki; Abo, Tatsuhiko; Aiba, Hiroji.

In: Genes to Cells, Vol. 4, No. 2, 1999, p. 87-97.

Research output: Contribution to journalArticle

@article{54ad1f50ea3847d6af381d6375f6c1cd,
title = "Regulation of intrinsic terminator by translation in Escherichia coli: Transcription termination at a distance downstream",
abstract = "Background: Rho-independent terminators in Escherichia coli are DNA sequences of 30-50 bp consisting of a GC-rich dyad symmetry sequence followed by a run of T residues in the nontemplate strand. The transcription termination at the Rho-independent terminator occurs within the T-tract in vitro. It has been believed that the transcription termination at the Rho- independent terminator occurs within the T-tract in vivo, as established in vitro, and therefore the 3' ends of mRNAs are mostly generated as a direct result of transcription termination. However, how the transcription termination occurs and how the 3' ends of mRNAs are formed in living cells remains to be studied. Results: We developed a double terminator system in which a second Rho-independent terminator was placed downstream of the crp terminator. This system made it possible to detect transcripts that pass through the crp terminator by Northern blotting. We found that most of the crp transcripts extend beyond the crp terminator. The transcriptional read- through at the crp terminator was reduced when the translation of crp mRNA was interrupted. The level of the read-through transcript decreased with distance between the two terminators, suggesting that transcription termination occurs at multiple positions beyond the crp terminator. Conclusion: We conclude that most RNA polymerase reads through the crp terminator in the natural situation and terminates transcription over a wide region downstream of the crp terminator, resulting in heterogeneous primary transcripts that are subsequently processed back to the terminator hairpin. We propose that ribosome translation to the crp stop codon causes read- through of the terminator. The regulatory effect of translation on Rho- independent termination may be a general phenomenon at other operons.",
author = "Hiroyuki Abe and Tatsuhiko Abo and Hiroji Aiba",
year = "1999",
language = "English",
volume = "4",
pages = "87--97",
journal = "Genes to Cells",
issn = "1356-9597",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Regulation of intrinsic terminator by translation in Escherichia coli

T2 - Transcription termination at a distance downstream

AU - Abe, Hiroyuki

AU - Abo, Tatsuhiko

AU - Aiba, Hiroji

PY - 1999

Y1 - 1999

N2 - Background: Rho-independent terminators in Escherichia coli are DNA sequences of 30-50 bp consisting of a GC-rich dyad symmetry sequence followed by a run of T residues in the nontemplate strand. The transcription termination at the Rho-independent terminator occurs within the T-tract in vitro. It has been believed that the transcription termination at the Rho- independent terminator occurs within the T-tract in vivo, as established in vitro, and therefore the 3' ends of mRNAs are mostly generated as a direct result of transcription termination. However, how the transcription termination occurs and how the 3' ends of mRNAs are formed in living cells remains to be studied. Results: We developed a double terminator system in which a second Rho-independent terminator was placed downstream of the crp terminator. This system made it possible to detect transcripts that pass through the crp terminator by Northern blotting. We found that most of the crp transcripts extend beyond the crp terminator. The transcriptional read- through at the crp terminator was reduced when the translation of crp mRNA was interrupted. The level of the read-through transcript decreased with distance between the two terminators, suggesting that transcription termination occurs at multiple positions beyond the crp terminator. Conclusion: We conclude that most RNA polymerase reads through the crp terminator in the natural situation and terminates transcription over a wide region downstream of the crp terminator, resulting in heterogeneous primary transcripts that are subsequently processed back to the terminator hairpin. We propose that ribosome translation to the crp stop codon causes read- through of the terminator. The regulatory effect of translation on Rho- independent termination may be a general phenomenon at other operons.

AB - Background: Rho-independent terminators in Escherichia coli are DNA sequences of 30-50 bp consisting of a GC-rich dyad symmetry sequence followed by a run of T residues in the nontemplate strand. The transcription termination at the Rho-independent terminator occurs within the T-tract in vitro. It has been believed that the transcription termination at the Rho- independent terminator occurs within the T-tract in vivo, as established in vitro, and therefore the 3' ends of mRNAs are mostly generated as a direct result of transcription termination. However, how the transcription termination occurs and how the 3' ends of mRNAs are formed in living cells remains to be studied. Results: We developed a double terminator system in which a second Rho-independent terminator was placed downstream of the crp terminator. This system made it possible to detect transcripts that pass through the crp terminator by Northern blotting. We found that most of the crp transcripts extend beyond the crp terminator. The transcriptional read- through at the crp terminator was reduced when the translation of crp mRNA was interrupted. The level of the read-through transcript decreased with distance between the two terminators, suggesting that transcription termination occurs at multiple positions beyond the crp terminator. Conclusion: We conclude that most RNA polymerase reads through the crp terminator in the natural situation and terminates transcription over a wide region downstream of the crp terminator, resulting in heterogeneous primary transcripts that are subsequently processed back to the terminator hairpin. We propose that ribosome translation to the crp stop codon causes read- through of the terminator. The regulatory effect of translation on Rho- independent termination may be a general phenomenon at other operons.

UR - http://www.scopus.com/inward/record.url?scp=0032903811&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032903811&partnerID=8YFLogxK

M3 - Article

VL - 4

SP - 87

EP - 97

JO - Genes to Cells

JF - Genes to Cells

SN - 1356-9597

IS - 2

ER -