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EXPRESSION
Gene
expression
Other
RNAs
Prokaryotes
STEP1
INITIATIONOFTRANSCRIPTION
Figure 8.11
signma protein recognize gene & bring pol, where transcription starts & goes to right D
- RNA pol
Prokaryotic
STEP2
ELONGATION
Prokaryotic
U
A
T
GACTTAGCGTGCATAGCA 3'
||||||||||||||||||
CTGAATCGCACGTATCGA 5'
A
A
T
G-G-G-C-C-C-G-A-C3'
| | | | | | | | |
C-C-C-G-G-G-C-T-G
G
A
complementary, antiparallel
elongation
5'
UU
UA
CC
AU
GU
UU
AA
A
GG
GC
C
CG
U
CU
AC
U
UA
A
CU
5' TTTACCATGTTTAAAGGGCCCGACTACTTAACT
|||||||||||||||||||||||||||||||||
3' AAATGGTACAAATTTCCCGGGCAGATGAATTGA
mRNA
STEP3
TERMINATION
Helicase breaks H bond
Prokaryotic
Translation
CU
UA
A
CU
GACTTAGCGTGCATAGCA 3'
||||||||||||||||||
CTGAATCGCACGTATCGA 5'
A
A
T
G-G-G-C-C-C-G-A-C3'
| | | | | | | | |
C-C-C-G-G-G-C-T-G
AA
AG
GG
CC
AU
GU
UU
ribosomes
UA
nascent
protein
CC
C
GU
C
UA
UU
The process of
reading the
genetic code in
the messenger
RNA (mRNA)
and using that
information to
synthesize the
polypeptide
(protein) that is
specified in the
mRNA.
A A G-G-G-C-C-C-G-A-C
5' TTTACCATGTTTAAAGGGCCCGACTACTTAACT
|||||||||||||||||||||||||||||||||
3' AAATGGTACAAATTTCCCGGGCAGATGAATTGA
Prokaryotic
!
mRNA
5'
PROKARYOTIC
5'
3'
3'
5'
Note the
direction of
information
flow.
Note
polarity
ribonucleotides
Transcription
mRNA
5'
3'
mRNA
5'
3'
amino acids
Translation
protein
protein
N = amino
protein
C = carboxy
protein
to make protein
N
N
3'
5'
Protein added
R added
R assemble
Genetic code
64 codons:
Thecodonsarereadthreenucleo?desinarowwithnogapsandwith
nopunctua?onbetweenthecodons
Met
Asn
Asn
Asn
Ala
no overlapping codons
Thecodeisdegenerate.
Thecodonsaredecodedby
tRNAintheconnesofa
ribosome.
Glu
Glu
Ala
Asp
tRNAs
tRNA single RNA chain folded into 2d cloverleaf
tRNA: small RNA molecules participate in translation of mRNA
not part of ribosome, have extensive 2nd structure
produced by INTRAmolecular base pairing btw nucleotides
-tRNA contains many highly modified &
unusual nucleotides
- following transcription, tRNA contains only
standard set of 4 nucleotides
- Unusual nucleotides are produced by
post-transcription enzymatic modifications
- tRNA has 4 stems & 3 loops (4th loop is
called Variable Loop) locate btw TUC stem
& anticodon stem
- D loop is named for modified nitrogenous base
found within it, dihydrouracil
- TUC loop is named for 4 invariant nucleotides
found within it (thymidylate, pseudouridylate,
cytidylate, guanylate)
Threonine
Th
re
on
in
Inosine
tRNA
Ribosomesarethesiteofpolypep?desynthesis.
Ribosomes
arecomplex
structures
composedof
RNAand
protein.
Fig. 8.23
8-38
Mechanismoftransla?on
IN PROKARYOTES
Ini?a?onsetsstageforpolypep?de
synthesis.
Firstaminoacidincorporatedinbacteria
Elonga?onduringwhichaminoacidsareaddedtogrowing
polypep?de add amino acid as ribosomes move 5' 3' d
Ribosomesmovein53direc?onrevealingcodons.
Addi?onofaminoacidstoCterminus
215aminoacidspersecond
Termina?onwhichhaltspolypep?desynthesis
Nonsensecodonrecognizedat3endofreadingframe
Releasefactorproteinsandhaltpolypep?desynthesis
nonsense=stop
Initiation
Fig. 8.25 a
Elongation
Termina?onoftransla?on
- Simultaneous synthesis of many copies of polypeptide from single mRNA
Fig. 8.25 c
Pleasereviewtheseclassesofmuta?on
Muta?onsinagenescodingsequencecanalterthegeneproduct.
Missensemuta?onsreplaceoneaminoacidwithanother.
Nonsensemuta?onschangeanaminoacidspecifyingcodontoastopcodon.
FrameshiTmuta?onsresultfromtheinser?onordele?onofnucleo?deswithin
thecodingsequence.
Silentmuta?onsdonotalteraminoacidspecied.
Genetic map
1. He wants to know the distance/how far apart 2 genes are/ same or dif
genes--> measure RF
5. The gene is co-linear the sequence of AA- begin of gene is begin of protein ??
3. He can detect
recombination bc 2 mutants
can cross over and give fully
functional Trp 8-7
Fig. 8.4
genetic map
He wants to know the distance/how far apart 2 genes are/ same or dif genes--> measure RF
Acodoniscomposedofmorethanone
nucleo?de.
Dierentpointmuta?onscanaectsameaminoacid.
Eachcodoncontainsmorethanonenucleo?de.
Eachnucleo?deispartofonlyasinglecodon.
Inferredbecauseeachpointmuta?onalteredonlyone
aminoacid.
He can detect recombination bc 2 mutants can cross over and give fully functional Trp Then he takes product of each mutant
genes --> protein --> sequence it
8-8
CrickandBrennerpropose
thattheeachwordis3
nucleo?deslongandtheyare
readcon?nuouslywithout
anypunctua?on. insertion & deletion
Thebeginningoftransla?on
establishesthereading
(forward & backward)
frame.
3 reading frames
Phenotypiceectof
frameshiTmuta?onsis
usuallyveryseverelossof
proteinfunc?on.
Fig. 8.6
Solve: homologous
recombinaton to put them
together
restore a frameshift
-----G--------------x
----------------A---------G---------A---insert G, remove A
Fig. 8.6
FC7
Original
mutation
Second
mutation
rIIB+ revertant
Fig. 8.5b
10
rIIB
Fig. 8.5c
FC0
FC7
rIIB
FC7
Fig. 8.5d
Phenotype
or
+
Mutant
or
++
Mutant
or
or
++++
or
+++++
Mutant
Wild type
or
or
+++
or
++++++
Wild type
12
What
does
each
codon
specify?
Phe Phe
Phe ...C
Analyze radioactive
polypeptides synthesized
N... Phe
Synthetic
mRNA
3'
Phe
Phe
Phe
Invitro translational
system plus radioactive
amino acids
Fig. 8.7
Keydiscoveries
ThemRNAandproteinarecolinear.The5endof
mRNAcorrespondstotheamino(N)terminusof
theproteinANDthe3endofthemRNA
correspondstothecarboxy(C)terminusofthe
protein.
Thegene?ccodeisdegenerate>20aminoacids
but61codonsforaminoacidsand3stop
codons.
Thegene?ccodeisuniversal.
invitrotransla?onexperiments
Summary
Codonconsistofatripletcodoneachofwhichspeciesanaminoacid.
Codeshowsa5to3direc?on.
Codonsarenonoverlapping.
Codeincludesthreestopcodons,UAA,UAG,andUGAthatterminate
transla?on.
Codeisdegenerate.
Fixedstar?ngpointestablishesareadingframe.
AUG
UAGinanini?a?oncodonwhichspeciesreadingframe.
53direc?onofmRNAcorrespondswithNterminustoCterminusof
polypep?de.
Muta?onmodifythemessageencodedinsequence
FrameshiTmuta?onschangereadingframe.
Missensemuta?onschangecodonofaminoacidtoanotheraminoacid.
Nonsensemuta?onschangeacodonforanaminoacidtoastopcodon.
8-15
Operon