Molecuar Dissection Of Relationship Between Autophagy And Apoptosis

In order to understand the relationship between apoptosis and autophagy, autophagy 5 was cloned from human fetal brain and B lymphocyte eDNA libraries by PCR, and examined gene structure using bioinformatics annotation. Through search in public database and sequence comparisons and assembly 4 related sequences, APG5 genomic sequence obtained was 144 159bp. By bioinformatics software, the structure of introns, exons, splicing sites, promoter and polyA were demonstrated. The hAPG5 gene had 8 exons, and a new isoform missing exon 3 was confirmed, which was hereby designated as human autophagy 513 (hAPG5 13; LOCUS AF293 841, Genbank). Moreover, both APG5/APG5 13 cDNAs in vitro were transfected by LIPOFACTIN for expression in L02 and Hela cells. Under confocal microscopy, both hAPG5 and Sf3 fusion proteins were visualized in cytosol of apoptosis cell and time of expression forward following apoptosis induced. Through phylogenetic analysis, APG5 was confirmed horizon transition.from single celled organism to multicellular organism and evolutionary conservation and APO5 is an important gene for life. Compression with APG5, protein prediction of APG513 show 4.3 %: extracellular, including cell wall and 4.3 %: Golgi distribution and lost 1 Protein kinase C, I Casein kinase II and I Tyrosine kinase phosphorylation site. Differential gene expression profiles between with and without 3-MA (inhibitor of autophagy) by 4096 cDNA microarray is described. The obtained data were analyzed. Among of 151 differential expression genes, 74 were up regulated; 77 down-regulated. APG5 gene remained no different. Electron microscopically, apoptosis morphology can be observed no matter 6 induced or inhibited autophagy.We conclude that although autophagy and apoptosis may be different ways but employ same regulation mechanism at special stage of apoptotic development or utilizing same protein in the maintenance of the integrity of apoptotic and autophagic bodies. We find apoptotic body formation have two types; one is by budding; other by scattering which was named by us. The latter means the apoptotic cells were separated by membrane cistern into several regions and then apoptotic bodies formed respectively and finally these bodies scattered simultaneously within the cell matrix. In present investigation the type of scattering was predominant in formation of apoptotic bodies. We believe this type is also predominant on the other cells apoptosis, because by scattering do not need more energy.