关键词: 染色质重组, PCR, 衰老, RNA, 基因敏感性

Abstract: Although the set of genes is virtually the same in all tissues,differential gene expression is appeared in cells of different kinds.Differentiation and ageing are associated with regulation of gene expression that is a fundamental mechanism in eukaryotic development and survival.The sensitivity to DNase I of actively transcribed genes seems to be a general phenomenon.The purpose of the study is to test whether RNAs obtained from different organs or cells can enhance susceptibility of albumin gene to DNase I digestion in BALB/c mouse brain chromatin assembled.RNAs extracted from rat liver,lung,kidney,brain,tRNA from yeast and synthesized RNAs (23nt completed with mouse alb gene) were added to a system of chromatin reconstitution that was achieved by dialysis from high ionic strength solution.Assembled chromatin was digested with DNaseⅠ(12.5μg/mL) at 20 ℃ for 1 min,then PCR assay was used to detect the level of albumin gene digested.PCR products (1200 bp) were run on a 6% polyacylamide gel and analyzed by silver stain assay.RNAs from different organs and synthesized RNAs all increased the sensitivity of albumin gene to DNaseⅠ attack in mouse assembled chromatin.The effect was more obvious in liver and lung RNAs than in kidney and brain ones.tRNA from yeast did not enhance the sensitivity of albumin gene to DNaseⅠ digestion.RNA increased albumin gene sensitivity to DNaseⅠ in a dose-dependent manner.We report here for the first time that RNAs can enhance susceptibility of albumin gene to DNaseⅠ digestion.The effect is associated with RNA sources or sequences.It is generally agreed that the formation of gene sensitivity to DNaseⅠ,by unfolding of a tightly packed chromatin fiber,is the first step in gene activation,then RNAs that recognize complementary DNA sequences may be the specific factors that affect DNA supercoiling and determine the sensitivity of gene to DNaseⅠ digestion.Here we describes "RNA Population Gene Activating Model" that gives a logical interpretation of events leading to expression of specific genes during normal development and differentiation,in the same time,explains ageing and oncogenesis.Gene expression in eukaryotic cells requires two level regulations.The first may be controlled by RNAs that locate complementary regions within the genomes and make these regions loosened potentially,and the second is mainly involved in sequence specific and nonspecific proteins by which genomic regions bound by RNAs are unfolded.In eukaryotic cells,RNA fragments cleaved from all transcripts mix together to form "RNA populations" in which the majority is intron RNA.Every type of RNA fragments and its homologous sequences act as a group to form certain concentration in which repetitive sequences are more effective.If it is considered that there are many groups of RNA fragments in a particular cell,then different groups of RNA fragments are presented in dissimilar cell types of differentiation.Between DNA replication and nucleosome formation,RNA fragments in nuclear liquid will compete with DNA for binding to complement regions,then the chromatin regions bound to RNA can not be wrapped to form typical nucleosomes.After DNA doubles and is divided into 2 cells,these regions containing atypical nucleosomes become loose by function of non-histone.Transcriptionally active regions of chromatin are loose conformation but loosened regions are not always transcriptionally active.In every division,cells surfer in the described procedure that genes express RNAs,then RNAs recognize and imprint DNA.There are different RNA populations in different cells so that they imprint different genes,which is the primary mechanism by which same genes have expression distinctness.Since loosened genes are similar to bacterial operator system,factors in environment around cells play roles in inducing different gene expression to form different RNA population,which is the primary reason of cell differentiation.RNA population produced by certain impressions in genome can not imprint to form the same ones,otherwise immortal cells will be emerged,so that this program also controls ageing and oncogenesis.