Only cytidines 5' of the apolipoprotein B mRNA mooring sequence are edited.
A 22 nucleotide 'cassette' containing three distinct sequence elements is required for efficient apolipoprotein B (apoB) RNA editing. One of the elements, the mooring sequence, has been proposed as the apoB RNA-recognition element in binding 66 and 44 kDa proteins during editosome assembly. We show here that editing can only take place on cytidines 5' of the mooring sequence; thereby providing further proof for the proposed orientation-specific assembly of editing factors at the editing site. We also provide direct evidence for the existence of a cryptic mooring sequence immediately downstream of the wild-type editing site (between nucleotides 6688-6710). When a cytidine is introduced 5' of this site (at nucleotide 6687), editing occurs at wild-type (nucleotide 6666) and cryptic sites. The cytidine to be edited is not involved in 66 and 44 kDa protein binding, and therefore, the data suggest that the cryptic mooring sequence binds these factors, and may facilitate factor recruitment to the wild-type editing site. These data may also explain why wild-type levels of editing on chimeric, RNA editing substrates can only be achieved when the 22 nucleotide editing cassette is flanked 3' by apoB RNA sequence.
Backus
J W
JW
Department of Biochemistry, University of Rochester, NY 14642.
Schock
D
D
Smith
H C
HC
D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D003562:Cytidine / Q000235:genetics*; D017931:DNA Primers; D015870:Gene Expression; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017393:RNA Editing; D012333:RNA, Messenger / Q000235:genetics*; D013329:Structure-Activity Relationship
D001055:Apolipoproteins B; D017931:DNA Primers; D012333:RNA, Messenger; D003562:Cytidine