In vitro reconstitution of apolipoprotein B RNA editing activity from recombinant APOBEC-1 and McArdle cell extracts.
Apolipoprotein B (apoB) RNA editing activity involves a site-specific cytidine to uridine transition catalyzed by a cytidine deaminase, APOBEC-1, in the context of a multi-protein-containing editosome. In the absence of yet to be characterized "auxiliary" proteins, APOBEC-1 lacks RNA editing capacity. Recombinant APOBEC-1 has been engineered to bind nickel resin and used in affinity chromatography of the auxiliary proteins from McArdle rat hepatoma cell extracts. We demonstrate activation of APOBEC-1 RNA editing activity under these conditions through the association of a subset of extract proteins having approximate molecular masses of 145, 87, 75, 66, 61, and 50 kDa and a heterogeneous grouping of 45- to 35-kDa proteins. These data suggest that the components of the editosome can be partially purified from extracts through APOBEC-1 affinity chromatography.
Yang
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Department of Biochemistry, University of Rochester, NY 14642, USA.
Smith
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HC
D000818:Animals; D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D002474:Cell-Free System; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D008969:Molecular Sequence Data; D017393:RNA Editing; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D001055:Apolipoproteins B; D017931:DNA Primers; D012333:RNA, Messenger; D016601:RNA-Binding Proteins