Characterization of a sequence (hlyR) which enhances synthesis and secretion of hemolysin in Escherichia coli (original) (raw)

Summary

A sequence (_hly_R) of about 600 bp which enhances the expression of hemolysin (HlyA) more than 50-fold was identified in the plasmid pHly152-specific hemolysin (hly) determinant. Deletion of this entire _hly_R sequence led to the same low level of hemolysin synthesis and excretion as that expressed by the recombinant plasmid pANN202-312. _Hly_R was active in cis but its activity was orientation-dependent. The enhancing sequence, _hly_R, is separated from the promoter _phly_I transcribing _hly_C, _hly_A and possibly hly_B by more than 1.5 kb including an IS_2 element. Stepwise removal of the _hly_R sequence from its 5′ end by exonuclease III (_Exo_III) digestion yielded several types of deletion mutants which expressed decreasing amounts of hemolysin. A similar observation was made when _hly_R was shortened by _Exo_III from its 3′ end, which suggests that more than one functional region may be present in the hly_R sequence. A deletion of 717 bp within the adjacent IS_2 element reduced the activity of hly_R only slightly, indicating that IS_2 is not directly involved in the enhancement mechanism but that it may support an optimal positioning in _hly_R relative to the hly promoter. The nucleotide sequence of _hly_R is rich in A+T and does not contain an extended open reading frame, but exhibits several sequence motives that may represent sites for protein binding and DNA bending.

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Authors and Affiliations

1. Institut für Genetik und Mikrobiologie der Universität Würzburg, D-8700, Würzburg, Federal Republic of Germany
   Monika Vogel, Jürgen Hess, Irene Then & Werner Goebel
2. Departamento de Microbiologia. Facultad de Biologia, Universidad de Barcelona, E-08071, Barcelona, Spain
   Antonio Juarez

Authors

1. Monika Vogel
2. Jürgen Hess
3. Irene Then
4. Antonio Juarez
5. Werner Goebel

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Communicated by E. Bautz

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Vogel, M., Hess, J., Then, I. et al. Characterization of a sequence (hlyR) which enhances synthesis and secretion of hemolysin in Escherichia coli.Molec. Gen. Genet. 212, 76–84 (1988). https://doi.org/10.1007/BF00322447

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• Received: 19 November 1987
• Issue date: April 1988
• DOI: https://doi.org/10.1007/BF00322447

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