第8章RNA转录后的加工讲解.pptVIP

和传统酶的区别： (1)一般的酶是纯的蛋白质，而核酶是RNA或带有蛋白的RNA； (2)核酶既是催化剂又是底物。而酶仅催化反应。 The first step in mRNA 3 end formation is the selection of the site within the pre-mRNA for processing.? Cleavage polyadenylation specificity factor (CPSF), a multi-subunit complex, binds at the AAUAAA polyadenylation signal found 10-30 bases upstream from the 3 end of most mRNAs, while CstF binds to a UG-rich sequence typically found 10-30 bases downstream from the 3 end.? The co-operative binding of these two complexes to the RNA and protein-protein interactions between them sets up the subsequent cleavage and polyadenylation reactions that generate the mRNA 3 end. This mechanism appears to be conserved between Human and Drosophila.? There are good Drosophila homologues for all of the subunits of both CPSF and CstF; mammalian polyadenylation sequences appear to function in Drosophila; AAUAAA and UG-rich sequences can be found at the 3 ends of most Drosophila genes. The protein encoded by su(f) is the largest subunit of CstF, and makes contact with the other CstF subunits, and with CPSF.? Alterations to the protein encoded by su(f) affect the CPSF-CstF interaction, and appear to affect selection of sites for mRNA 3 end formation.? In su(f) mutants, changes have been described in forked, ADH-related and su(f) mRNAs such that sites distant from the promoter are used in preference to sites closer to the promoter. We have cloned all of the subunits of both CstF and CPSF and are using Drosophila genetics and other approaches (such as yeast two-hybrid assays) to study the interactions between the subunits of CstF, between CPSF and CstF, and between these complexes and other components of the transcriptional and splicing apparatus.? We have recently identified lethal mutations in the 73 kilodalton subunit of CPSF thought to encode the actual endonuclease responsible for the cleavage.? We have also shown that mutations in genes for the largest subunit of CPSF and the largest subunit of RNA polymerase II affec