title: Molecular modelling of the biosynthesis of the RNA-editing enzyme APOBEC-1, responsible for generating the alternative forms of apolipoprotein B.
abstract: We discovered in 1987 that the shorter form of apolipoprotein B (B48) synthesized in the intestine is due to the action, previously unrecognized in mammalian cells, of an mRNA-editing process, and more recently we demonstrated that this was due to a specific enzyme (APOBEC-1) with cytidine deaminase activity. We show here, by sequence alignment, molecular modelling and mutagenesis, that APOBEC-1 is a cytidine deaminase, responsible for editing apoB mRNA, and that is related in crystal structure to the cytidine deaminase of Escherichia coli (ECCDA). The two enzymes are both homodimers with composite active sites formed with loops from each monomer. In the sequence of APOBEC-1, three gaps compared with ECCDA match the size and contour of the minimal RNA substrate. We propose a model in which the asymmetric binding of one active site to the substrate cytidine which is positioned by the downstream binding of the product uridine and that this helps to target the other active site for deamination.
authors.affiliation: National Heart & Lung Institute, Imperial College School of Medicine, Hammersmith Hospital, London. j.scott@ic.ac.uk
authors.forename: J
authors.initials: J
authors.lastname: Scott
authors.forename: N
authors.initials: N
authors.lastname: Navaratnam
authors.forename: C
authors.initials: C
authors.lastname: Carter
mesh_terms: D000071479:APOBEC-1 Deaminase; D000818:Animals; D001055:Apolipoproteins B / Q000096:biosynthesis*; D001161:Arteriosclerosis / Q000378:metabolism*; D001483:Base Sequence; D003564:Cytidine Deaminase / Q000096:biosynthesis* / Q000235:genetics; D004252:DNA Mutational Analysis; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D017393:RNA Editing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
chemical_list: D001055:Apolipoproteins B; D012333:RNA, Messenger; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D000071479:APOBEC-1 Deaminase; C000605089:APOBEC1 protein, human; D003564:Cytidine Deaminase