Lau P P, Zhu H J, Nakamuta M, Chan L
Departments of Cell Biology and Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
J Biol Chem. 1997 Jan 17;272(3):1452-5. doi: 10.1074/jbc.272.3.1452.
Apolipoprotein (apo)B mRNA editing is mediated by a multiprotein editosome complex. Apobec-1 is the catalytic component of this complex, but other proteins involved in editing have not been identified. We used the yeast two-hybrid system to identify an apobec-1-interacting protein, ABBP-1. ABBP-1 contains 331 amino acid residues and is identical to a previously reported human type A/B hnRNP except for a 47-residue insertion at its C-terminal region. It contains typical RNP motifs at its N-terminal half and glycine-rich motifs in the C-terminal region. Northern blot analysis indicates that ABBP-1 mRNA is distributed in multiple human tissues. By deletion analysis, we mapped the apobec-1-binding region to the glycine-rich domain. ABBP-1 also binds to apoB mRNA transcripts around the editing site and can be UV-cross-linked to them in vitro. Immnodepletion of ABBP-1 from an active apoB mRNA editing tissue extract inhibits its editing activity. Down-regulation of ABBP-1 in an apobec-1-expressing HepG2 cell line by transfection with an antisense ABBP-1 cDNA construct leads to inhibition of endogenous apoB mRNA editing. We conclude that ABBP-1 is an apobec-1-interacting protein that may play an important role in apoB mRNA editing.
载脂蛋白(apo)B信使核糖核酸(mRNA)编辑由一种多蛋白编辑体复合物介导。载脂蛋白B mRNA编辑酶催化多肽1(Apobec-1)是该复合物的催化成分,但参与编辑的其他蛋白质尚未得到鉴定。我们利用酵母双杂交系统鉴定出一种与Apobec-1相互作用的蛋白质,即载脂蛋白B mRNA编辑辅助蛋白1(ABBP-1)。ABBP-1含有331个氨基酸残基,除了在其C末端区域有一个47个残基的插入外,与先前报道的人类A/B型不均一核糖核蛋白(hnRNP)相同。它在其N末端一半含有典型的核糖核蛋白基序,在C末端区域含有富含甘氨酸的基序。Northern印迹分析表明,ABBP-1 mRNA分布于多种人体组织中。通过缺失分析,我们将Apobec-1结合区域定位到富含甘氨酸的结构域。ABBP-1还能结合编辑位点周围的apoB mRNA转录本,并能在体外与它们进行紫外线交联。从活性apoB mRNA编辑组织提取物中免疫去除ABBP-1会抑制其编辑活性。通过用反义ABBP-1 cDNA构建体转染在表达Apobec-1的肝癌细胞系(HepG2)中下调ABBP-1会导致内源性apoB mRNA编辑受到抑制。我们得出结论,ABBP-1是一种与Apobec-1相互作用的蛋白质,可能在apoB mRNA编辑中起重要作用。