title: Protein splicing of the Saccharomyces cerevisiae VMA intein without the endonuclease motifs.
abstract: The protein splicing element (intein) of the vacuolar ATPase subunit (VMA) of Saccharomyces cerevisiae catalyzes both protein splicing and site-specific DNA cleavage. It has been demonstrated that the conserved splice junction residues are directly involved in protein splicing and the central dodecapeptide motifs are required for DNA cleavage. To examine whether the splicing activity of the intein can be structurally separated from the endonuclease motifs, we made large in-frame deletions at the central region of the intein. We demonstrate for the first time that protein splicing can proceed efficiently after the removal of the central region of the intein including the endonuclease motifs. Our results suggest that the N- and C-terminal regions of the Sce VMA intein may form a separate domain that is not only catalytically sufficient for protein splicing but also structurally independent from the endonuclease domain.
authors.affiliation: New England Biolabs, Inc., Beverly, Massachusetts 01915, USA.
authors.forename: S
authors.initials: S
authors.lastname: Chong
authors.forename: M Q
authors.initials: MQ
authors.lastname: Xu
mesh_terms: D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002686:Chitin / Q000235:genetics / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism*; D016368:Frameshift Mutation; D058619:Maltose-Binding Proteins; D008970:Molecular Weight; D019154:Protein Splicing; D006180:Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D017384:Sequence Deletion; D025262:Vacuolar Proton-Translocating ATPases
chemical_list: D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D002686:Chitin; D004720:Endonucleases; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases