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Nucleic Acids Research Advance Access originally published online on August 18, 2006
Nucleic Acids Research 2006 34(15):4258-4268; doi:10.1093/nar/gkl574
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Nucleic Acids Research, 2006, Vol. 34, No. 15 4258-4268
© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

The gene of an archaeal {alpha}-L-fucosidase is expressed by translational frameshifting

Beatrice Cobucci-Ponzano1, Fiorella Conte1, Dario Benelli2, Paola Londei2, Angela Flagiello3, Maria Monti3, Piero Pucci3, Mosè Rossi1,4 and Marco Moracci1,*

1 Institute of Protein Biochemistry—Consiglio Nazionale delle Ricerche Via P. Castellino 111, 80131 Naples, Italy 2 Dipartimento di Biochimica Medica e Biologia Medica (DIBIM), Università di Bari—Piazzale Giulio Cesare 70124 Bari, Italy 3 CEINGE Biotecnologie Avanzate s.c.a r.l., Dipartimento di Chimica Organica e Biochimica Università di Napoli Federico II, Complesso Universitario di Monte S. Angelo, via Cinthia 4, 80126 Napoli, Italy 4 Dipartimento di Biologia Strutturale e Funzionale, Università di Napoli ‘Federico II’ Complesso Universitario di Monte S. Angelo, Via Cinthia 4, 80126 Naples, Italy

*To whom correspondence should be addressed. Tel: +39 081 6132271; Fax: +39 081 6132277; Email: m.moracci{at}ibp.cnr.it

Received April 19, 2006. Revised July 21, 2006. Accepted July 22, 2006.

The standard rules of genetic translational decoding are altered in specific genes by different events that are globally termed recoding. In Archaea recoding has been unequivocally determined so far only for termination codon readthrough events. We study here the mechanism of expression of a gene encoding for a {alpha}-L-fucosidase from the archaeon Sulfolobus solfataricus (fucA1), which is split in two open reading frames separated by a –1 frameshifting. The expression in Escherichia coli of the wild-type split gene led to the production by frameshifting of full-length polypeptides with an efficiency of 5%. Mutations in the regulatory site where the shift takes place demonstrate that the expression in vivo occurs in a programmed way. Further, we identify a full-length product of fucA1 in S.solfataricus extracts, which translate this gene in vitro by following programmed –1 frameshifting. This is the first experimental demonstration that this kind of recoding is present in Archaea.


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