An open reading frame in intron seven of the sea urchin DNA-methyltransferase gene codes for a functional AP1 endonuclease

Author:

CIOFFI Anna Valentina1,FERRARA Diana2,CUBELLIS Maria Vittoria3,ANIELLO Francesco4,CORRADO Marcella1,LIGUORI Francesca1,AMOROSO Alessandro1,FUCCI Laura4,BRANNO Margherita1

Affiliation:

1. Biochemistry and Molecular Biology Laboratory, Stazione Zoologica ‘A. Dohrn’, Villa Comunale 80121 Naples, Italy

2. Dipartimento Medicina Sperimentale, Sezione di Fisiologia Umana e Funzioni Biologiche, Integrate ‘F. Bottazzi’, Second University of Naples, Via Costantinopoli 16, 80138 Naples, Italy

3. Department of Organic and Biological Chemistry, University of Naples ‘Federico II’, Via Mezzocannone 16, 80138 Naples, Italy

4. Department of Genetics, General and Molecular Biology, University of Naples ‘Federico II’, Via Mezzocannone 8, 80138 Naples, Italy

Abstract

Analysis of the genome structure of the Paracentrotus lividus (sea urchin) DNA methyltransferase (DNA MTase) gene showed the presence of an open reading frame, named METEX, in intron 7 of the gene. METEX expression is developmentally regulated, showing no correlation with DNA MTase expression. In fact, DNA MTase transcripts are present at high concentrations in the early developmental stages, while METEX is expressed at late stages of development. Two METEX cDNA clones (Met1 and Met2) that are different in the 3′ end have been isolated in a cDNA library screening. The putative translated protein from Met2 cDNA clone showed similarity with Escherichia coli endonuclease III on the basis of sequence and predictive three-dimensional structure. The protein, overexpressed in E. coli and purified, had functional properties similar to the endonuclease specific for apurinic/apyrimidinic (AP) sites on the basis of the lyase activity. Therefore the open reading frame, present in intron 7 of the P. lividus DNA MTase gene, codes for a functional AP endonuclease designated SuAP1.

Publisher

Portland Press Ltd.

Subject

Cell Biology,Molecular Biology,Biochemistry

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