Reflections on protein splicing: structures, functions and mechanisms.
Twenty years ago, evidence that one gene produces two enzymes via protein splicing emerged from structural and expression studies of the VMA1 gene in Saccharomyces cerevisiae. VMA1 consists of a single open reading frame and contains two independent genetic information for Vma1p (a catalytic 70-kDa subunit of the vacuolar H(+)-ATPase) and VDE (a 50-kDa DNA endonuclease) as an in-frame spliced insert in the gene. Protein splicing is a posttranslational cellular process, in which an intervening polypeptide termed as the VMA1 intein is self-catalytically excised out from a nascent 120-kDa VMA1 precursor and two flanking polypeptides of the N- and C-exteins are ligated to produce the mature Vma1p. Subsequent studies have demonstrated that protein splicing is not unique to the VMA1 precursor and there are many operons in nature, which implement genetic information editing at protein level. To elucidate its structure-directed chemical mechanisms, a series of biochemical and crystal structural studies has been carried out with the use of various VMA1 recombinants. This article summarizes a VDE-mediated self-catalytic mechanism for protein splicing that is triggered and terminated solely via thiazolidine intermediates with tetrahedral configurations formed within the splicing sites where proton ingress and egress are driven by balanced protonation and deprotonation.
Anraku
Yasuhiro
Y
Graduate School of Science, University of Tokyo, Tokyo, Japan. anraku@jcom.home.ne.jp
Satow
Yoshinori
Y
D047669:Exteins / Q000235:genetics / Q000502:physiology; D047668:Inteins / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019154:Protein Splicing / Q000235:genetics / Q000502:physiology*; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D053778:Thiazolidines / Q000737:chemistry / Q000378:metabolism
D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; D053778:Thiazolidines; D006180:Proton-Translocating ATPases; C081687:VMA1 protein, S cerevisiae