The gene of an archaeal alpha-L-fucosidase is expressed by translational frameshifting.
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.
Cobucci-Ponzano
Beatrice
B
Institute of Protein Biochemistry-Consiglio Nazionale delle Ricerche, Via P. Castellino 111, 80131 Naples, Italy.
Conte
Fiorella
F
Benelli
Dario
D
Londei
Paola
P
Flagiello
Angela
A
Monti
Maria
M
Pucci
Piero
P
Rossi
Mosè
M
Moracci
Marco
M
D001105:Archaea / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D016368:Frameshift Mutation; D018965:Frameshifting, Ribosomal; D019848:Gene Expression Regulation, Archaeal / Q000502:physiology*; D048229:Sulfolobus solfataricus / Q000235:genetics; D005644:alpha-L-Fucosidase / Q000235:genetics*
D005644:alpha-L-Fucosidase