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An Escherichia coli cis-acting antiterminator sequence: The dnaG nut site

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Summary

The Escherichia coli rpsU-dnaG-rpoD operon contains an internal transcription terminator T1 located in the intergenic region between the rpsU and dnaG genes (Smiley et al. 1982). By cloning T1 as a small 127 bp fragment into the terminator probe plasmid pDR720 between the trp operator promoter and the assayable galK gene, it was shown that T1 acts as a strong transcription terminator, comparable in strength to the 3′ operon terminator T2. However, an operon sequence that occurs 5′ to T1 within the coding region of the rpsU gene and which has homology with the lambda nut site, (Lupski et al. 1983) when placed 5′ to T1 in the pDR720 plasmid construct, modifies transcription through T1 allowing expression of the galK gene. This sequence, called the dnaG nut site also modifies the termination activity of the external operon terminator T2. It is proposed that the dnaG nut site is a cis-acting element of an antitermination system in E. coli.

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Abbreviations

nut :

N gene protein utilisation site

CAT:

chloramphenicol acetyl transferase

CoA:

co-enzyme A

RF:

replicative form DNA

DTNB:

5,5′-dithiobis-(2-nitrobenzoic acid)

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Author notes

  1. N. Almond

    Present address: Imperial Cancer Research Fund Laboratory, Lincoln's Inn Fields, P.O. Box No. 123, WC2A 3PX, London, England

Authors and Affiliations

  1. Biochemistry Department, NYU Medical Center, 550 First Avenue, 10016, New York, NY, USA

    N. Almond, V. Yajnik, P. Svec & G. N. Godson

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  1. N. Almond
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  2. V. Yajnik
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  3. P. Svec
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  4. G. N. Godson
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Communicated by R. Devoret

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Almond, N., Yajnik, V., Svec, P. et al. An Escherichia coli cis-acting antiterminator sequence: The dnaG nut site. Mol Gen Genet 216, 195–203 (1989). https://doi.org/10.1007/BF00334356

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  • DOI: https://doi.org/10.1007/BF00334356

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