title: Protein trans-splicing and functional mini-inteins of a cyanobacterial dnaB intein.
abstract: A 429 aa theoretical intein is encoded in the dnaB gene (DNA helicase) of the cyanobacterium Synechocystis sp. strain PCC6803. This intein is shown to be capable of protein splicing with or without its native exteins when tested in E. coli cells. A centrally located 275 amino acid sequence (residues 107-381) of this intein can be deleted without loss of the protein splicing activity, resulting in a functional mini-intein of 154 aa in size. Efficient in vivo protein trans-splicing was observed when this mini-intein was split into a 106 aa N-terminal fragment containing intein motifs A and B, and a 48 aa C-terminal fragment containing intein motifs F and G. These results indicate that the N- and C-terminal regions of the Ssp DnaB intein, whether covalently linked with each other or not, can come together through non-covalent interaction to form a protein splicing domain that is functionally sufficient and structurally independent from the centrally located endonuclease domain of the intein.
authors.affiliation: Biochemistry Department, Dalhousie University, Halifax, Nova Scotia B3H 4H7, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Wu
authors.forename: M Q
authors.initials: MQ
authors.lastname: Xu
authors.forename: X Q
authors.initials: XQ
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry*; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000201:enzymology*; D004265:DNA Helicases / Q000737:chemistry*; D053488:DnaB Helicases; D004720:Endonucleases / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D019154:Protein Splicing / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004720:Endonucleases; D004265:DNA Helicases; D053488:DnaB Helicases