MOLECULAR BIOLOGY MCDB 3500
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This 16 page Class Notes was uploaded by Mrs. Willis Mante on Thursday October 29, 2015. The Class Notes belongs to MCDB 3500 at University of Colorado at Boulder taught by Staff in Fall. Since its upload, it has received 28 views. For similar materials see /class/231845/mcdb-3500-university-of-colorado-at-boulder in Molecular, Cellular And Developmental Biology at University of Colorado at Boulder.
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Date Created: 10/29/15
Chapter 10 Eukaryotic RNA polymerases and their promoters Page 276295 Downstream promoter element Upstream BRE TATA an DPE element 25 I Initiator Initiator PyPyANTAPyPy is suf cient by itself to direct transcription Downstream promoter elements DPE quite common in fruit y genes Found 30 bp downstream of the transcription initiation site Have the consensus sequence GATCG behave as a TATAbox BRE GCGCGACGCC TFIIB binding site Upstream element GC box GGGCGG CCGCCC Binds Spl transcription activator cat box CCAAT Binds CTF transcription activator Class I promoter Pol I promoter Relatively simple one template the rRNA precursor gene Copyright The McGrawHm Companies Inc Permissron required for reproduction or display 100 D V O CO to co N o o n v v a n l m lt1 ltr lt1 N n 2 I m Ix ID V CO l V lt 0 o o 39 39 39 5 3 75 co lt1 co N no to no lt co m m N 397 397 T P 9 3 lt0 N 0 Relative TXN ef ciency 01 O 25 I 156 107 45 1 20 gt Source Hepnmed 1mm Learned H M TK Learned M M Hammer and H T Tnan Isl46848 VQBB Cupynght 1986 wwn perrmssm at ElsewerSclencel 1 Core element Upstream promoter element The spacing between the two elements is important Figure 1033 200 460 120 80 3940 1 20 I I I I I I I I I I I I ela ve promoter strength Wild Iype A4 Lsae73 100 L5 95 73 P 40 L3 9557 4i 40 LSSBMS g 20 Lees45 Io 1 340745 i 0 LS10734 lt5 LsIowsm 100 LS54 94 Il 301 A1 La 4 n 4 35 28 LSsws I 1004 3 LS 137 181 30 Class III promoter Pol III promoter Cupytht T V W WWW w dyspmy l Further 3 deletion I Z I I I I I No transcription mm L zgn munv pr a 5mm 3 any Class III promoter Pol III promoter Flgu re 1 Saginaw Classwal class III promoter 53 rRNA Bo Box C IRNA or M VA RNA Box A Box B Unlike class I and II promoters classical class III promoters locate Within the genes U6 snRNA 7SL and 7SK RNA gene still need elements 5 of the genes Their promoters resemble class II promoters nonclassical class III promoter Enhancers and Silencers Figure 1038 210 kb Elements that are not a part of the promoter but can either enhance enhancer or inhibit silencer transcription at a manner that is position and orientationindependent How do enhancers work 1 An enhancers binds transcription factors activators to activate transcription 2 An enhancer can localize upstream Within or downstream of a gene How do enhancers work 1 An enhancers binds transcription factors activators to activate transcription 2 An enhancer can localize upstream Within or downstream of a gene Cupyngmr39 4 nc my A1 180 bp Com5M Tquot 5 739mv 1 WW quot 39i quot394 quot39 39quot 1quot quot gtJ Cu 71 m9quot N Am bp uuunnuuuwuvpmy 12345678910111213 st x2 A B C D B r Aw Hr g g 123456789101112M A X X 7 0 X3lt X2 2 3 2quot D 1 C 5 Yzb nunO X2 X3 00 c 39 C aI I 7wa1 A1 A2 b n m quotQ mms Chapter 11 General Transcription Factors in Eukaryotes Page 303309 Polymerases need help to nd the promoters The factors that attract polymerases to their respective promoters are called general transcription factors General transcription factors combine with an RNA polymerase to form an open promoter complex or a preinnitiation complex to activate transcription Class II factors work with Pol II Class II preinitiation complex Six class II factors TFIIA TFIIB TFIID TFIIE TFIIF and TFIIH Preinitiation complex is assembled in a precise order D gt DA gt DAB gtDABPolF gt DABPolFEH Two important assays to examine proteinDNA interaction DNase I footprint analysis Gel mobility shift assay or Gel retardation assay D N A fragment Prutein incubate with protein Protein beam to spewi c DNA quttence lDigcst wnh DNase ill chnatum II UM tit Radioartive label l Divide sample Incubate without protein DigPsl with DNase itiiiijiit EIECU OIJI39IUI ESiS Aumrnciiugraphy Regier of DNA plolL Cli d by protein binding Wi i i1iiiiky D i i DNA mm xixmmme Divide sample Protein Incubate with Incubate without protein protein Protein lmunrl 0 DNA we mmmmm Electrophoresis Amoracl lugrnphy DNAeproiei n complex i Free DNA
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