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EK Bio1 Ch 2 (cont..

Regulation of transcription
o In prokaryotes
mRNA typically includes several genes in a single
transcript
Jacob-Monod model
Operon = operator + promoter + genes
contributing to single prokaryotic mRNA
o Genes outside of operon may code for
activators/repressors
Lac operon in e. coli
o Codes for enzymes that allow e. coli to
import & metabolize lactose when
glucose is nor present in sufficient
quantities
o Activated when glucose is scarce and
lactose is present
o Mechanism: (38-39)
Low glucose levels -> high cAMP
levels
cAMP activates CAP
CAP binds to CAP site adjacent to
promoter on lac operon
Positive control: CAP activates
promotor
Gene repression: when lactose not
present, lack repressor protein
binds to operator, prevents
expression of lac genes
o In eukaryotes
mRNA includes only 1 gene per transcript
modification of RNA
o post-transcriptional modifications offur in both eukaryotic
and prokaryotic cells
eukaryotes: only in nucleus
bacterial genome does not contain introns
o purpose of modifications:
help molecules that initiate translation to recognize
the mRNA
protect mRNA from degradation
eliminate extraneous sequences of nucleotides from
the transcript before translation

provide mechanism for variability in protein products


produced by a single transcript
o primary transcript = first nucleotide sequence arrived at
through transcription
much longer than mRNA that will be translated into
protein
portions of transcript (introns) are spliced out
before exiting nucleus
exons are joined end-to-end
**introns stay IN nucleus, exons Exit nucleus to
be translated
snRNPs = small nuclear ribonucleoproeins, act
as ribozymes
o spliceosome = complex of snRNP +
associate proteins
o 5 end of eukaryotic transcription is capped via GTP before
transcription is finished
5 cap serves as attachment site in protein synthesis
& as protection against enzyme degradation (via
exonucleases)
3 end has long series of adenine nucleotides (poly A
trail)
o alternative splicing in eukaryotic cells
allows cell to incorporate different coding sequences
into mature mRNA
can create variety of mRNA molecules for translation
from single DNA coding seq
human genome is 20,000 25,000 coding regions,
but codes for >100,000 protein products
introns help determine possible splicing patterns &
promote protein production
introns tend to be highly conserved between
species
translation: nucleotide sequence of mRNA to amino acid seq of
corresponding protein
o triplet code: 3 nucleotides (codon) to code for 1 amino acid
degenerative >1 series of 3 nucleotides can code
for same AA
**Stop codons: UAA, UGA, UAG
**Start codon: AUG
Number of possible combinations of any three
nucleotides: 43 = 64
o Types of RNA

mRNA = template that carries genetic code from


nucleus to cytosol in form of codons
tRNA
two ends:
o one contains anticodon, series of 3
nucleotides that binds to complementary
codon sequence on RNA
o other carries AA that corresponds to
codon, to be added to growing
polypeptide chain
some flexibility in bonding at the third-bp position
wobble pairing
all translation uses ribosomes
small subunit + large subunit
prokaryotic: 30s + 50s = 70s
eukaryotic: 40s + 60s = 80s
the nucleoleus manufactures ribosomes (euk only)
mechanism: (p 43)
initiation
after post-transcriptional processing (euk),
mRNA leaves nucleus though nuclear pores,
enters cytosol
initiation factors help attach 5 end to small
ribosomal subunit
tRNA containing 5-CAU-3 gathers methionine
and settles into p-site
this signals large subunit to join & form
initiation complex
elongation
ribsosome slides down mRNA one strand at a time 5>3 while matching each codon to complimentary
tRNA anticodon
corresponging AAs attached to tRNA are bound
together into growing polypeptide
elongation requires energy
when tRNA attaches to P-site, new tRNA attaches to
neighboring A site, peptide bond forms between the
two via peptidyl transferase activity
tRNA with methionine moves to E-site, and then exits
ribosome, tRNA carrying new dipeptide moces to p
side, a site is open for next until stop codon reaches
p
termination

when stop codon reaches A site, release factor


proteins add to end of polypeptide chain, which frees
it from tRNA and ribosome
ribosome breaks into subunits to be used later
o polypeptide begins folding during translation, assisted by
chaperons
post-translational modification
o regulate gene expression by affecting which translational
products become proteins
o may add sugars, lipids, or phosphate groups, or cleave the
polypeptide
o final destination = related to location of translation
o 20 AA seq called signal peptide near front of polypeptide is
recognized by protein called RNA signal-recognition particle
(SRP) that carries entire ribosome complex to receptor
protein on ER
protein grows across membrane, then either released
into lumen or remains partially attached to ER
signal peptide removed by enzyme
signal peptides may also be attached to polypeptides
to target them to mitochondria, nucleus, or other
organelles
DNA Replication: Mitosis
o Semiconservative: each DNA copy contains one strand of
original DNA
o Mechanism governed by group of proteins called replisome
o Replication begins in middle of chromosome, at the origin
of replication
Single origin on prok, multiple on euk.
Bidirectional process, two replisomes proceed in
opposite directions

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