6.1. 6.2. RNA6.3. RNA6.4. 6.5. 6.6. 6.7. 6.8. 6.9.

(transcription):DNADNA RNA4rNTPATP CTPGTPUTPRNA RNARNA Double Strand DNA ()

(sense strand) [ coding strand]: DNA (antisense strand) [ template srand] : RNA (60) AU GC

DNARNA AUCG RNA RNA53DNA 35 RNA 53 (DNA ,RNA )

RNA promotion. elongation. terminaton (promoter)(terminator) (transcription unit) () polycistron in operon, monocistron, No operon 1

RNA

(RNA Transcription promotion in prokaryots)

: (operon):

(promoter)DNARNA () RNA polymerase (E. coli)1 (Holo Enzyme)(Core Enzyme)

(1) Holo Enzyme DNA ( ) DNA() 1014mol 107mol 1

2 Core Enzyme DNA DNA DNA 1011mol 60

2 1 RNApol Holo Enzyme Sextama Box(35), 2 2 2 23

E.coli 70325428 11 2 RNApol DNA DNA DNA

3 NMP Editing Rho RNA 3end

E siteelongation site RifR:NTP Editing 4 RNAsense strand SSB Holoenzyme I siteinitiation site . Rifs: ATPGTP ATPGTP

DNA DNA/RNA DNA DNA

E. coli RNA polymerase36.5 KD36.5 KD151 KD155 KD11 KD70 KD

promoter Prok.E.coli 1 1 CAP-cAMP CAPcatabolite gene Activator Protein cAMP 2 RNApol 35 ~ 10 R B

2 RNA 1 Sextama Sextama Box -35RNA RNA R T82T84G78A65C54A45 : RNApol

2 Pribonow Pribonow Box -10RNA B T80A95T45A60A50T96 TATPUAT 4 13TATA Box

3 initiation site1 RNA NTPATPGTP

a. R

b RNApol10 ATPribnow 1217bp 10 RNApol DNA

1217bp

c. RNApolIE +1CAT,T 6~9 4 DNA

1217bp

3 CAP-cAMP CAP I lac

I

: E.coli CAP ATP cAMP cAMPCAP, RNApol cAMPCAPcAMP CAPRNApol RNApol

,,, lac1CAP 70 40 70 50 IR 50 40 cooperativity

2 CAPcAMPRNApol Sextama Box,10 CAPcAMP Sextama Box GCGC Pribnow Box

:1RNApol2 RNApol

4RNApol DNA DNase ----------40bp footprinting RNApolDNA DNA DNAKlenow3, 5 DNA

DNA

DNARNApol DNase DNase

RNApol DNA +20 ~ 5040 60 ~ 70 bp .. 5 135 10 35 TTGACA 10 TATAAT

a. b. c. : 35 10

2 3510 17bp 17bp 17bp

E.Coli

RNApol 1 RNApol - RNApol RNApol RNApol 2 RNApol rRNA5.8S18S 28S

RNApol hnRNAsnRNA hnRNAmRNA RNA heterogeneous nuclear RNA snRNA RNA small nulear RNA) RNApol tRNA5S rRNAAlu snRNA RNApol

3 500KDa, 7~12 RNApol C (carboxy terminal domain CTD) CTD TyrSerProThrSerProSer C

CTD Ser Thr RNApol A RNApol B CTD B A RNApol 10 RNApol

1 RNApol , 1 cap site Prok.CAT 2 TATAHogness Goldberg-Hogness 30 T82A97T93A85A63T37A83A50T37

Pribnow TATA3 CAATCAAT box 75bp GGC/TCAATCT G ()

4 enhancer far upstream sequence 100 SV40DR -107~178 179 ~ 250 72bp TATA

: 72bp 72bp 5 GC GC box 90 GGGCGG

6 octamer element OCT ATTTGCAT KB GGGACTTTCC ATF GTGACGT 7 SV406GC

,

2 RNApol 1 a 5SRNA tRNA b snRNA

2 5S rRNA 5S rRNA 5580 5580

5S rRNA RNApol55bp3 AC 5S rRNA AB tRNAVARNA RNApol

:1RNApolDNA23RNApol 4RNApol5RNApol

RNApol (), DNA,RNApol

RNApol

RNApol

RNApol TFA TFB TBPBRTB TFC 5

TBP RNApol TBP RNApol,TBPSL1 RNApol RNApolTBPTATA TBP , positioning factor

DNA 1Prok. DNA , , 2 Euk.

Prok RNA NTPRNA 3-OH NTPRNA

1 RNApol DNA17bp DNA RNAase______10bp

2 RNA RNApol .. RNApol ..DNA DNA RNA-DNA

3 1 RNA 30 ~ 50 , 15AAsec 2 DNAGC GC 8~10 RNA: ?

RNA terminator RNA RNA Prok.Euk.

1 1 2

2 : . 7~20 bpIRG/C . 6 ~ 8 U

GC

3 1 IR GC 2

UG/C

1 1 RNA RNApol 60 2 RNApol 6 ~ 8U RNApol RNADNA rUdA RNA

RNADNA RNApol 3 U4 IRU IRGC U5 DNAUAT AT

2 1 read through RNApol IR RNApol 2 a NTPase

b RNA50ntRNANTPase RNA3 aRNARNA5 bRNA53NTP c RNApol

RNApol RNApol

4 RNA DNA RNA RNApol AU

5 Prok. Prok.

* ------ * T7------RNApol *

PMPLPRPR

PNPQ

(PL nutLtR1nutR nutPN utilization)nutRNApol RNApol,

:12 3

3 1 RNApolProk. RNApol SV40: U 2 RNApol U U

Prok.mRNA rRNAtRNA 5

rRNA 1 rRNA 5S16S23StRNA rrnE.coli rrn tRNA

rRNA 16S rDNA tDNA.23S rDNA.5S rDNA tDNARNAse

tRNA 1tRNA 1 tRNAtRNA rRNA 2 tRNA 3CCA 3CCA

3 CCA Prok. tRNA CCA Phage Euk.

2 tRNA 1 RNAaseP tRNA 5 tRNA 2 RNAaseD tRNACCA a RNAaseP RNAaseD b RNAaseQRNAaseYRNAaseP3

3 tRNAtRNA nucleotidyl transferase ATPCTP tRNA3CCA a E.coli CCA b tRNA4 RNAase RNAaseORNAaseP2

3E.coli tRNATyr1 350base

tRNATyr1

RNA m5C m6A CH3H3C Prok. Euk.

tRNA

Euk Pre-RNA processing in Eukaryotes

rRNA rRNA5.8S18S28SrDNA47S45S a b c45SRNA

: 18S rRNA5 18S rRNA5.8S rRNA28S rRNA 5.8S rRNA 28S rRNA Euk. 5S rRNA

mRNA-------- 1 0Cap-0 m7GpppXpYp---------- m7G N7 1 Cap-1 m7GpppXmpYp---------- 2-O A N6 2Cap-2 m7GpppXmpYmp 2-O AGCU

HNCH3m7G0

: Cap0 Cap1 Cap2 2 SSAM RNA-----capping enzyme

3 1 ------RNA Cap0 Cap1,2 2 mRNA 5 3 RNA Cap1 Cap2 Euk.mRNAm6A

mRNA-------- A 1 3-----200bp (Euk.mRNA) poly(A)+ poly(A)- RNA3 poly(A) E.coli poly(A)+1962 2 poly(A) a RNApoly(A) ATP

+ nATP Mg++ Mn++ (A)n + nPPi b (360KDa) poly(A) poly(A) 1320bp AAUAAA GUGUGUG Euk.

:1Prok. rRNA tRNA2Euk.rRNA

3 poly(A) c poly(A) mRNA poly(A) 1 2 mRNA 3 a b poly(A) mRNA poly(A)

4 poly(A) a oligo (dT) RNA mRNAb dToligo (dT) cDNA

1 split geneRNA RNARNA intronRNA RNARNA DNA excon

RNARNA splicing RNA 5 3

2 1982 Davies central core structure: 4 10 ~ 20bp4

group : group : group : GUAG mRNA tRNA tRNA

3 mRNA RNA tRNA

tRNA 1 2tRNA3 Nt40400 14~16bp tRNA 3

tRNAphe

4 ..tRNA tRNAtRNAtRNA()

RNA 5OH 33 23 OH

a3OH 3 OH 2 b 5 2 tRNA

RNAtRNA

1 1 5 3 -------U G

5 exonUintronGexon3

2 a 5 PQRS3 10~12bp PQRS

b interal guide sequence IGS #

2 rRNA 1 rRNA35S26SrRNA 5 U G 3 rRNA rRNA26S 2 35SRNA GTP GDP GMP 3 G414base 5

4 G G 3-OH 5 G- 3OH

3-OH 3 ,RNA

5 -------- . ,G () :

Cech

:1pol(A) 2Euk.tRNA

3 35SRNA414base 1

2 C15: UUU 5 : RNA 3 L19 Cech L19 polyC

L19

L19

4 a 35SRNA --- RNARNA rearrangease RNARNA isomerase

bL19 polyC L19 RNA---- ribozyme R

1 7 Branch Site 3 6bp ~ 12bp

Branch Site 10-50Nt IR,(A IR, 5--GUGCG--------------------PyPuPyPyUAPy-----PyAU3

2Splicing mechanism

mRNA 1 Euk.100 ~ 300 RNA, 105~106 snRNA snRNP scRNA scRNP 2mRNA

Breathnach-Chambon rule GUAG

3Splicing mehanism ) SnRNA (or ScRNA) , ( spliceosome) Spliceosome SnRNAU1U2U5 U4U6 U1 5 U2 A U1U2 5 3 U1 3 U1U2mRNA U4U5U6

lariat exons spliceosome lariat

Discontinuoustranscription and Trans-splicing

RNA cis-splicing RNAtrans-spling 1982 Vander Ploeg mRNA 535 bases mRNA 35bases leader sequence

1 200 copies 3100base mRNA mRNA

5 GU mRNA mRNA 3 AG, A

mRNA mRNA 1986 W. J. Marphy R. E. Sutton 1 3 mRNA 5 2 Y

RNA 1RNApol2E.coliProk.3Prok.45Prok.6E.coli

7 8 tRNA9 tRNA 10poly(A)11Prok.12Euk.mRNA13rRNA

1516 14mRNA3poly(A)

AUCG RNADNA (RNADNA)

Proximal--- distal DNA107mol R,B site 1014mol RNA

rifamycin rifampinIstreptollydiginE

Consensus ,RNApol

RNA

DNase 40bpfootprinting

DNA RNApol RNApol

17bp,DNA12 ~ 17bp

DEAERANTyr Thr B A10A GC TATATATACAAT G ,72bpH2BOTC2CAAT1TATAsnRNA

5SrRNADNA RNApol

RNADNA

17Transcription: the synthesis of a single-stranded RNA from a double-stranded DNA templateDNA

15AAsec RNA

RNA 3OHRNARNA

6 ~ 8 U

RNApolb RNApol 1:10 ,c RNA50ntRNANTPaseRNApol

PmRNApolRNARNAse

tRNA 5()RNAaseD tRNACCACCAa E.coli CCA b ATPCTPtRNA c tRNA

45S11045SRNAEuk.5S rRNA tRNA5S rRNA

Klug,RNARNA4 Euk.mRNAm6AmRNA,poly(A)PABPPABPSepharosetRNA340400, 14~16bp-----------tRNA3

tRNA5OH3tRNAtRNA

c

a G3OH 5U 35 b 5 3OH 3G

3 L19 Cech L19 polyC

mRNA

U1U2 5 3U1 3A 2 OH 5G 3-OH3G lariat exons

2001986 W. J. Marphy R. E. Sutton