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Etty Widayanti, Ssi. Mbiotech
Etty Widayanti, Ssi. Mbiotech
Nov 2009
B-galactosidase
H20 (energy)
galactose + glucose
1. Constitutive
2. Regulated / inductive
Lac Operon
lac genes
promoter
Z
operator
cAMP is low
Lac Z
Remember lactose
galactose + glucose
If lactose & glucose are present - no lacZ is made until
glucose is depleted.
How?
High cAMP is necessary for activation of lac operon
cAMP is bound by CAP (catabolite activator protein)
cAMP-CAP binds to distal part of promoter and facilitates
transcription
Operons
General Theme:
A metabolite controls the expression of a battery of
genes that have evolved to utilize it (CATABOLISM)
Other Examples:
trp - tryptophan biosynthesis
ara - arabinose utilization
his - histidine biosynthesis
Think About:
Genetic logic of negative or positive control
i. e. repression & activation
Tryptophan Operon
1. trpE is first gene in operon
2. trpE mRNA has long leader (untranslated 5' region)
3. region of mRNA works as attenuator:
in presence of tryptophan, transcription is halted
about 140 bases into mRNA
in absence of tryptophan, transcription continues
Mechanism is complex, but logical
leader sequence contains short 11 amino acid peptide with
two trp residues
when trp is abundant, trp-tRNA can be used to translate
leader mRNA which terminates transcription
when trp is limiting, translation stalls and transcription is
permitted
Trp Attenuation
Transcription Control
Post-Transcriptional Control
Translational Control
Post-Tranlational Control
DNA
A. Transcription
B. mRNA processing
C. mRNA transport
mRNA
cytoplasm
D. translation
polypeptide chain
E. Protein processing
active protein
nucleus
A.Transcription
Binding of transcription factors to
special sequences in DNA slows or
speeds transcription.
Chemical modifications and chromosome
duplications affect RNA polymerases
physical access to genes.
B. mRNA processing
New mRNA cannot leave the nucleus
before being modified, so controls over
mRNA processing affect the timing of
transcription.
Controls over alternative splicing
influence the final form of the protein.
C. mRNA transport
RNA cannot pass through a nuclear pore
unless bound to certain proteins.
Transport protein binding affects where
the transcript will be delivered in the
cell.
D. Translation
An mRNAs stability influences how long it is
translated.
Proteins that attach to ribosomes or initiation
factors can inhibit translation.
Double-stranded RNA triggers degradation of
complementary mRNA.
E. Protein processing
A new protein molecule may become
activated or disabled by enzymemediated modifications, such as
phosphorylation or cleavage.
Controls over these enzymes influence
may other cell activities.
Transcriptional Control
Transcription start site
Transcriptional Control
The basal promoter
The basal promoter contains a sequence of 7 bases (TATAAAA) called
the TATA box (this is very similar to the -10 box or Pribnow box found
in prokaryotes) .
It can be bound by Transcription Factor IID (TFIID read TF2D)
which is a complex of some 10 different proteins including
- TATA-binding protein (TBP), which recognizes and binds to the
TATA box
- other protein factors which bind to TBP - and each other - but
not
to the DNA.
The basal or core promoter is found in all protein-encoding genes. This
is in sharp contrast to the upstream promoter whose structure and
associated binding factors differ from gene to gene (i.e. they are unique
to each specific gene).
REFERENCES
Campbell, N.A., Reece, J.B. and Mitchell, L.G. 2004.
Biologi. Jilid ke-3. Ed ke-5. Penerbit Erlangga,
Jakarta.
Lehninger, A.L. 1982. Dasar-dasar biokimia. Jilid 3.
Terj dari Principles of biochemistry, oleh
Thenawidjaya, M. Penerbit Erlangga, Jakarta.
Starr, C., Taggart, R., Evers, C. and Starr, L. 2009.
Cell biology and genetics. Biology: The unity and
diversity of life. 12th ed. Brooks/Cole, Belmont.
Yuwono, T. 2005. Biologi molekular. Penerbit Erlangga,
Jakarta.