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Nucleic Acids Research, 1993, Vol. 21, No. 18 4200-4205
© 1993

MOLECULAR BIOLOGY

Mutations dissociating the inhibitory activity of the pokeweed antiviral portein on eukaryote translation and Escherichia coli growth

Jean-Michel Dore, Evelyne Gras, Florence Depierre and John Wijdenes

Innotheèrapie Laboratoires, 1 Boulevard A. Fleming BP 1937, 25020 Besancon Cedex, France

Received June 21, 1993. Revised August 9, 1993. Accepted August 9, 1993.

The pokeweed antiviral protein Is a rlbosome inactivating protein acting on eukaryotlc as well as on prokaryotlc ribosomes thus Is toxic for both cell types. Using the PCR technique to clone the PAP open reading frame, we characterized two cDNAs coding for proteins Inhibiting eukaryotlc translation process and which are not toxic for Escherichia coll, unlike the wild type protein. The sequence of the two cDNAs showed that the proteins contain only one and two point mutations. This result suggest that the wild type amlno acids In the mutated positions participate In the prokaryotlc ribosome recognition. These mutants might be useful for the construction of immunotoxins containing the pokeweed antiviral protein as toxin.


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