The Impact Of Alternative Splicing With 8 Amino Acids Of A1CF On Nuclear Location And A1CF Antibody Preparation

RNA editing is a very critical process of genetic adaption, including nucleoside indel and base replacement, which has a great impact on gene expression, producing different amino acids and new open reading frames to generate new gene products and take charge of various genetic functions.RNA editing is widely distributed in a variety of creatures, such as protozoon, mammals, plants, fungi, insects, viruses and so forth. Alcf is primarily discovered in the process of apoB mRNA editing. It contains three RNA recognition motifs(RRMs) and an ACF nuclear localization sequence. As a complementary effect factor of APOBEC-1, an editing complex can be formed by Alcf and APOBEC-1 in participation of apoB mRNA editing. Alternative splicing of A1CF can generate many mutants, one of which is 8 amino acids loss in nuclear localization sequence, but the impact of those 8 amino acids on the function of Alcf is still unknown.Hippo-Yap and Hedgehog signaling discovered in fruit fly are highly conserved in many species. Organ size, cell proliferation and apoptosis, cancer occurrence, tissue repair and regeneration are regulated by Hippo-Yap signaling. While, Hedgehog signaling plays an indispensable role in embryo development and a plenty of cellular biological functions, such as cell growth, survival, apoptosis and organ morphogenesis. In this article, alcf has been analyzed by many bioimformatic tools. Sequence alignment of amino acids, establishment of cladogram and protein structural domain analysis indicate that alcf is highly conserved in all the species. The mutant of alcf contains 8 amino acids deletion is also highly conserved in different species.The structural analysis of alcf illustrates that those 8 amino acids are located in the nuclear localization sequence of alcf. Thus, a fusion protein comprising alcf and GFP is constructed. The result implies that the deletion of those 8 amino acids would affect the alcf entrance to nuclear. Dual luciferase analysis demonstrates that A1CF(A 8aa), the mutant of alcf, can activate Hippo-Yap and Hedgehog signaling. Furthermore, we have prepared the polyclonal antibody of alcf in order to help the further investigation of alcf. In summary, all those results give us a hint that not only alcf but also Hippo-Yap and Hedgehog signaling act as very critical roles in cell growth and survival, and Hippo-Yap and Hedgehog signaling are most likely regulated by alcf in this whole process.