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expA: A conditional mutation affecting the expression of a group of exported proteins in Escherichia coli K-12

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Summary

A mutant of Escherichia coli K-12 was isolated as conditionally deficient in the expression of two exported proteins simultaneously (i.e. two acid phosphatases) The mutant was found to be thermosensitive on minimal medium at 37°C and above, but grew normally on rich media at these temperatures. The mutation, named expA and located at 22 min on the recalibrated linkage map, depressed the levels of six periplasmic enzymatic activities in bacteria grown at 37°C. At least ten proteins were greatly reduced in the periplasm under these conditions. The mutation also affected some outer membrane proteins, among which were the ompF protein and a protein which may be protein III, but had little effect on cytoplasmic membrane proteins. The gel patterns of the soluble cytoplasmic proteins were not modified except for one major protein of MW 47,000. The activities of β-galactosidase and of aspartate transcarbamylase were unmodified. After growth at 30°C no difference was observed between expA and expA + isogenic strains. The results are discussed with respect to the mechanism of protein export.

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Abbreviations

E.C. 3.1.3.2:

Acid phosphatase (optimum pH 2.5)

E.C. 3.1.3.2:

Acid phosphatase (optimum pH 4.5)

E.C. 3.1.3.1:

Alkaline phosphatase

E.C. 3.4.11:

Aminopeptidase-N

E.C. 2.1.3.2:

Aspartate transcarbamylase

E.C. 3.2.1.23:

β-galactosidase

E.C. 3.1.27.1:

RNase I

PNP-OH:

Para-nitro-phenol

PNA:

Para-nitro-anilide

PNP-P:

Para-nitro-phenyl-phosphate

Bis-PNP-P:

Bis-para-nitro-phenyl-phosphate

IPTG:

Isopropyl-β-thiogalactopyranoside, Nitrosoguanidine N-methyl, N'-nitro, N, Nitrosoguanidine, Tris Tris (hydromethyl) aminomethane

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Authors and Affiliations

  1. Service de Biochimie, Département de Biologie, Centre d'Études Nucléaires de Saclay, B.P. 2, F-91190, Gif-sur-Yvette, France

    Elie Dassa & Paul-Louis Boquet

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  1. Elie Dassa
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  2. Paul-Louis Boquet
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Communicated by H. Saedler

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Dassa, E., Boquet, PL. expA: A conditional mutation affecting the expression of a group of exported proteins in Escherichia coli K-12. Molec. Gen. Genet. 181, 192–200 (1981). https://doi.org/10.1007/BF00268426

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  • DOI: https://doi.org/10.1007/BF00268426

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