医用分子遗传学转录和转录铁后修饰.pptVIP

BMB400, 3_RNA processing_04 李 希 分子医学教育部重点实验室 lixi@shmu.edu.cn Post-transcriptional Processing of RNA Making ends of RNA RNA splicing Primary Transcript Primary Transcript-the initial molecule of RNA produced--- hnRNA （heterogenous nuclear RNA ） In prokaryotes, DNA →RNA →protein in cytoplasm concurrently In eukaryotes nuclear RNA Cp RNA Types of RNA processing A) Cutting and trimming to generate ends: rRNA, tRNA and mRNA B) Covalent modification: Add a cap and a polyA tail to mRNA Add a methyl group to 2’-OH of ribose in mRNA and rRNA Extensive changes of bases in tRNA C) Splicing pre-rRNA, pre-mRNA, pre-tRNA by different mechanisms. 5’ and 3’ ends of eukaryotic mRNA Capping of pre-mRNAs Cap=modified guanine nucleotide Capping= first mRNA processing event - occurs during transcription CTD recruits capping enzyme as soon as it is phosphorylated Pre-mRNA modified with 7-methyl-guanosine triphosphate (cap) when RNA is only 25-30 bp long Cap structure is recognized by CBC（cap-binding complex） stablize the transcript prevent degradation by exonucleases stimulate splicing and processing Functions of 5’ cap and 3’ polyA Need 5’ cap for efficient translation: Eukaryotic translation initiation factor 4 (eIF4) recognizes and binds to the cap as part of initiation. Both cap and polyA contribute to stability of mRNA: Most mRNAs without a cap or polyA are degraded rapidly. Shortening of the polyA tail and decapping are part of one pathway for RNA degradation in yeast. mRNA Half-life ? t? seconds if seldom needed ? t? several cell generations (i.e. ~48-72 h) for houskeeping gene ? ≈avg 3 h in eukaryotes ? ≈avg 1.5 min in bacteria Split gene and mRNA splicing Exon and Intron Exon is any segment of an interrupted gene that is represented in the mature RNA product. Intron is a segment of DNA that is transcribed, but removed from within the transcript by splicing together the sequences (exons) on either side of it. The sequence of steps in the production of matu