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Ways to Regulate Transcription Alternate sigma factor usage: controls selective transcription of entire sets of genes vegetative (principal s) s70 צמיחה heat shock nitrogen starvation +1 (16-19 bp) TTGACA s32 s60 TATAAT (5-9 bp) A +1 CNCTTGA (13-15 bp) CCCATNT (5-9 bp) A +1 CTGGNA (6 bp) TTGCA (5-9 bp) A !19.6.2008 – התרגיל ביום חמישי זה טרנסקריפציה באוקריוטים Pre-mRNA * microRNAs (miRNA) RNA molecules of about 21–23 nucleotides in length, which regulate gene expression. are processed from primary transcripts known as pri-miRNA. 5’UTR 3’UTR בתאי כבד בעכבר RNA polymerase מיקום 1+ יעילות ותדירות רמה נמוכה (בזלית) אינטרונים E S RE אין אחידות במיקום וברצפי הבקרה בין גנים שונים. שלבי הטרנסקריפציה Initiation • Elongation • Termination • קומפלקס פרה-איניציציה יציבות ותדירות יצירת קומפלקס זה קובעת את כמות וקצב סינטזת הRNA- אופן אקטיבציה של טרנסקריפציה בתאים אוקריוטים Stages in Initiation of Transcription • Bacterial transcription • Closed complex: holoenzyme+promoter • Open complex (DNA melting, not need ATP) • Abortive transcripton • Productive initiation – Transcribe past +9 – Sigma dissociates • Elongation • Eukaryotic transcription • Preinitiation complex (PIC) assembly • PIC activation (DNA melting, needs ATP) • Abortive transcription • Productive initiation – CTD phosphorylated – Promoter clearance • Elongation initiation Elongation NTPs TERMINATION • RNA polymerase meets the terminator • Terminator sequence: AAUAAA • RNA polymerase releases from DNA • Prokaryotes-releases at termination signal • Eukaryotes-releases 10-35 base pairs after termination signal קומפלקס האלונגציה transcription Transcription regulation Gene expression Gene expression Basics of cell biology:development Fertilized egg Basics of cell biology:development Cells differentiation is due to different gene Switches going on and off These cells are different because they express a subset of their 24,000 genes Promoter Gene To specify a new cell: place it in the right environment or EGF Hedgehog Wnt Promoter Gene Retinoic acid FGF HGF To specify a new cell: place it in the right environment or Substitute drugs/proteins for environment Mophogens Chromosome localization in interphase In interphase, chromosomes appear to be localized to a sub-region of the nucleus.