Synthesis of RNA molecules: Transcription. Hajnalka Horváth 2016
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1 Synthesis of RNA molecules: Transcription Hajnalka Horváth 2016
2 GENERAL FEATURES
3 The flow of genetic information (central dogma) The genetic information is transcribed from DNA to RNA Synthesis of RNA molecules using DNA as a template Primary transcript/pre-rna processing mature RNA
4 At the DNA containing part of the cells DNA serves as a template Enzyme: RNA polymerase (forms 3-5 phosphodiester bonds) The direction of the synthesis is 5-3 from the point of view from the new RNA molecule!!! (but from the point of view of DNA it is 3 to 5 ) Substrate: ribonucleoside-triphosphate β (beta) and γ (gamma) phosphate groups are released during the synthesis Bases in RNA: A, U, G, C (no T)
5 Transcription unit 1. Initiator/promoter region 2. RNA coding region 3. Terminator Asymmetrical No need for primers
6 Steps of transcription Initiation Transcriptosome binds here (enzymes for DNA denaturation, regulatory proteins, RNA polymerase etc.) Regulatory role of promoter Elongation Starts from +1 site RNA polymerase incorporates the complementary ribonucleotides (phosphodiester bond formation) 5 tryphosphate end of the newly synthesized RNA molecule The growing 3 end is attached to the active strand of the DNA Termination By different mechanisms in pro- and eukaryotes
7 TRANSCRIPTION IN PROKARYOTES
8 In the nucleoid region Coupled transcription, translation chromosome-polysome complex No nuclear envelope No processing in case of prokaryotic mrna The direction of transcription and translation are the same
9 Promoter in prokaryotes -35 and -10 (Pribnow-box) sequence RNA polymerase binds here Core enzyme + sigma subunit (the sigma subunit of RNA polymerase recognizes the promoter region) DNA denaturation happens here
10 Termination in prokaryotes Rho dependent ρ factor binds to a specific sequence on the RNA and cuts off the RNA molecule from the DNA by its helicase activity Rho independent the new RNA forms a hairpin by selfcomplementer basepairing and tears off the RNA molecule from the DNA (poly-u region after the hairpin forming region)
11 Pre-RNA processing in prokaryotes Posttranscriptional (the whole new RNA strand is already synthesized when the processing starts) Prokaryotic mrna is mature (no processing is needed) remember the chromosome-polysome complex Prokaryotic trna and rrna is processed by exo- and endonucleases
12 A little help B6NFIZU A very detailed description: gy/biochemistry-ii/rna-andtranscription/transcription-in-prokaryotes
13 TRANSCRIPTION IN EUKARYOTES
14 In the nucleus (chromatin=proteins+dna) DNA accessibility influences transcription (the proteins can overlie the transcription unit) Different RNA polymerases: No coupled transcription, translation Nuclear envelope mrna processing Mature RNA transported to the cytosol
15 Promoter in eukaryotes Transcription factors! (RNA polymerase cannot bind directly to the promoter ) Much more complex, more precise regulation is possible Core promoter: Right before the RNA coding region Sequences: TATA-box and InR Enhancer elements: Can be far away from the RNA coding region Influence the transcription
16
17 Elongation in eukaryotes Similar to the prokaryotic mechanism Elongation factors and other proteins are attached to the RNA polymerase
18 Geoffrey M. Cooper, Robert E. Hausmann-The Cell: A molecular approach
19 Termination in eukaryotes Polyadenilation sequence transcribed to the mrna Protein complex binds to this sequence and cuts the RNA strand (endonuclease) A2_03-11.html
20 Pre-RNA processing in eukaryotes rrna and trna: exo- and endonucleases, chemical modifications, addition of bases posttranscriptionally Cloverleaf structure of trna
21 Pre-RNA processing in eukaryotes mrna: 5 cap formation (protection against degradation, ribosome binding) 3 poly-a tail (protection against degradation, poly-a polymerase synthesizes it) splicing
22 Pre-RNA processing in eukaryotes Splicing: Eukaryotic pre-mrna contains exons and introns Performed by RNA-protein complexes (RNP)
23 A little help again vdbfhlo ce-4og4
24 Thank You for Your attention! mas_tree_rrna.html rrna transcription: the so called Christmas tree structure
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