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Molecular Biology Chapter 7
Transcriptional control of Gene Expression
88
Biology
Graduate
11/09/2012

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Term
Transcriptions role in gene expression
Definition
repression and activation of genes to produce specific gene products
Term
Repression means that
Definition
proteins are produced at low rates
Term
promoter
Definition
DNA sequences to which RNA polymerase and sigma factors bind to begin transcription
Term
How is the strength of a promoter regulated?
Definition
Consensus sequence has the most commonly occurring base at each of the positions in the -35 and -10 regions
Term
What is meant by the term “strength” of a promoter
Definition
Optimal binding site for RNA polymerase and sigma factor
Term
role of the sigma factor (sigma) in the initiation of transcription in E. coli
Definition
Sigma factor binds to RNA polymerase and to promoter DNA sequences, bringing the RNA polymerase enzyme to a promoter
Term
Where does sigma factor bind?
Definition
promoter region
Term
What enzyme does sigma factor associate with?
Definition
RNA polymerase
Term
What is the most common sigma factor
Definition
sigma 70
Term
What types of transcription is sigma factor involved with
Definition
Operon transcription, bacterial transcription
Term
What is an operon
Definition
Clustered genes which encode enzymes involved in a particular metabolic pathway or proteins that interact to form one multi-subunit protein
Term
Lac operon mechanism
Definition
Lac mRNA is repressed when E.coli is in an environment where there is no lactose. When there is glucose and lactose, E.coli prefers to metabolize glucose. When lactose is the only sugar available to metabolize E.coli represses transcription of the lac operon until lactose is present and this allows synthesis of only low levels of lac mRNA until the cytosolic concentration of glucose galls to low levels
Lac operon encodes three proteins required for the metabolism of lactose
Term
Operator
Definition
specific DNA sequence in the lac transcription-control region, controlled by lac repressor. When no lactose is present, the lac repressor binds to the lac operator, and the operon cannot be transcribed. When lactose is present, it binds to the lac repressor which causes it to dissociate from the lac operator which allows for operon transcription
Term
repressor
Definition
tetrameric protein which controls the lac operator, it is bound to operator when lactose is not present. When lactose is present, it binds to the repressor, which causes it to dissociate from the operator.
Term
lactose
Definition
sugar present in milk, a low levels repressor it bound to operator, a high levels it binds to repressor allowing for dissociation and gene transcription.
Term
glucose
Definition
a sugar favorably used for metabolism by E.coli. When it is in high concentrations, the repressor binds the lac operon. When the concentration falls E.coli synthesizes cAMP which binds to CAP protein and interacts with the polymerase bound to the promoter which stimulates the rate of transcription initiation
Term
CAP
Definition
catabolite activator protein, controls lac operon transcription under low glucose levels, dimeric protein, bound by cAMP when glucose levels are low
Term
cAMP
Definition
synthesized by E.coli when glucose levels are low in order to bind to CAP and interact with polymerase bound to the promoter and increase lac operon transcription initiation
Term
Enhancer
Definition
activators whose binding sites are 80-160 base pairs upstream from the start site, regulates transcription of genes by RNA polymerase containing sigma 54
Term
Function of Sigma 54
Definition
Sigma 54 associates with enhancers as activators which can be far away from the start site
Term
Two-component regulatory system
Definition
o Depending on two proteins for control, they control many responses of bacteria to changes in their environment
Term
example of a two-component regulatory system
Definition
E.coli glnA gene
Term
Factors, which regulate transcription elongation in E. coli
Definition
cytoplasm concentration of tryptophan
amino-acylated tRNA concentration
Term
transcriptional control in Prokaryotes
Definition
gene control serves mainly to allow a single cell to adjust to changes in its environment so that its growth and division can be optimized
Term
transcriptional control in eukaryotes
Definition
Primary purpose of gene control in eukaryotes is the execution of precise developmental decisions so that proper genes are expressed in the proper cells during embryonic development and cellular differentiation
Term
transcriptional control in both eukaryotes and prokaryotes
Definition
Transcriptional control is the primary means of regulating gene expression
Term
three different types of RNA polymerases found in eukaryotes
Definition
RNA polymerase I- transcribes pre rRNA
RNA polymerase II- transcribes mRNA, snRNAs, siRNAs, and miRNAs
RNA polymerase III- transcribes tRNAs, 5s rRNA, snRNA U6, and 7S RNA
Term
CTD function
Definition
CTD of RNA polymerase II is the carboxyl-terminal domain, heptapeptide repeat with a consensus sequence. It is critical for viability in yeast
Term
How is CTD modified
Definition
It becomes phosphorylated during transcription initiation and remains phosphorylated as the enzyme transcribes the template
Term
TATA box
Definition
conserved sequence in highly transcribed genes, found ~ 26-31 base pairs upstream of the transcription start site, acts similarly to an E.coli promoter to position RNA pol II for transcription initiation
Term
initiators
Definition
alternative promoter element (to TATA box), contain a cytosine at the -1 position and an adenine reside at the transcription start site (+1), nucleotide sequence directly surrounding the start site determines the strength of the initiator
Term
BRE
Definition
transcription factor recognition element
Term
IHR
Definition
initiator
Term
DPE
Definition
downstream core promoter element
Term
CpG islands
Definition
promoter regions for genes that encode proteins which are not required in large numbers, occur relatively rarely in genomes of mammals, CG sequences
Term
TBP
Definition
first protein to bind to a TATA box promoter, domain folds into a saddle shape, interacts with the minor groove of the DNA, and bends the helix
Term
TFIIB
Definition
monomeric protein, smaller than TFB, C-terminal domain makes contact with TBP and DNA on either side of the TATA box, N-terminal domain is inserted into the RNA exit channel of RNA pol II, N- terminal assists Pol II in melting the DNA strands at the transcription site and interacts with the template strand near the Pol II active site
Term
TFIIF
Definition
performed complex, binds with Pol II in positioning the polymerase over the start site
Term
TFIIE
Definition
creates a docking site for TFIIH
Term
TFIIH
Definition
completes assembly of the transcription pre-initiation complex
Term
Helicase Activity of TFIIH
Definition
uses energy from ATP to help unwind the DNA at the start site allowing Pol II to form an open complex in which DNA around the start site is melted and the template strand id bound at the polymerase active site
Term
Duel Role of TFIIH
Definition
Completes assembly of transcription pre-initiation complex and has Helicase activity
Term
Mutation of TFIIH
Definition
When a transcribing RNA pol becomes stalled, TFIIH recognizes the stalled pol and the associates with other proteins to repair the damaged DNA region. In mutations, repair of genes is inactive, and affected individuals have extreme skin sensitivity to sunlight and have incidence of cancer
Term
how enhancer sequences can regulate transcription of specific genes at a distance
Definition
Stimulate transcription from a promoter far away
Term
Relationship between activators and their domains and enhacers and their domains
Definition
Activators use activation domains
Enhancers use repressors
Term
How does DNAase footprinting reveal where a transcription factor bind
Definition
Takes advantage of the fact that when a protein is bound to a region of DNA, it protects that DNA sequence from digestion by nucleases. The appearance of a footprint (region protected by the bound protein that appears as a gap) indicates the presence of a transcription factor that binds that control element in the protein sample being assayed
Term
Constitutive gene expression
Definition
Constitutive regulated gene expression is unregulated, abnormally high expression due to mutation. Results from inactivation of a repressor that normally inhibits the transcription of those genes
Term
Mutation of repressors and activators
Definition
Mutation of repressor binding site = increased expression of a reporter gene
Mutation of an activator binding site = decreased expression of the linked reporter gene
Term
typical characteristics of the DNA binding domains of activators
Definition
have two domains
Term
typical characteristics of the DNA binding domains of repressors
Definition
can inhibit transcription from a gene they do not morally regulate when their cognate binding sites are placed close or far from the gene’s start site. Two functional domains: DNA binding domain and repressor domain
Term
typical characteristics of DNA binding domains of repressors and activators
Definition
Both domains continue to function when fused to another type of DNA-binding domain. Both domains function by interacting with other proteins
Term
Homeodomain
Definition
conserved 60-residue DNA-binding motif, present in eukaryotic transcription factors that function during development, similar to helix-turn-helix, found in genes that have master-control functions in human development
Term
example of homeodomain
Definition
development genes of drosophila
Term
Zinc-finger
Definition
regions that fold around a central zinc ion, producing a compact domain from a relatively short length of the polypeptide chain, common in DNA binding and transcription factors
Term
Leucine zipper domains
Definition
DNA binding domains, mutagenesis of leucine was required for dimerization
Term
Example of leucine zipper domains
Definition
DNA-binding domain of yeast GCN4 transcription factor
Term
Basic Helix-Loop-Helix (bHLH) proteins
Definition
DNA-binding domains of another class of dimeric transcription factors, form heterodimers
Term
modular nature of transcription factors in terms of DNA binding and RNA polymerase activation domains
Definition
Structural motifs bind specific DNA sequences, results in a noncovalent interaction between atoms in an alpha helix in the DNA-binding domain and atoms on the edge of the bases within the major groove in the DNA
Term
how chromatin remodeling influences transcription regulation
Definition
Activators or repressors exert their effects by binding to multi-subunit co-activators or co-repressors that influence assembly of Pol II transcription pre-initiation complexes by modulating chromatin
Term
What proteins play a role in the process of remodeling
Definition
Histones tail (H3 and H4)
Term
How does post-translational modification of chromatin proteins lead to regulation of transcription
Definition
Post-translational modification can aceylate Histones to mediate chromatin repression
Term
mechanism by which steroid hormone receptors regulate transcription
Definition
Induced conformational changes that modify their interactions with other proteins
Steroid hormones are homodimeric nuclear receptors, when bound to their ligands they can translocate to the nucleus and activate transcription of target genes
Term
five members of the nuclear receptor superfamily
Definition
Estrogen receptor
Progesterone receptor
Glucocoticoid receptor
Thyroxine receptor
Retinoic acid receptor
Term
Epigenic
Definition
inherited changes in the phenotype of a cell that do not result from changes in DNA sequence
Term
mechanisms observed in epigenetic regulation of transcription
Definition
Repression or activation that is maintained after cells replicate as the result of DNA methylation and/or post-translation modification of Histones, especially histone methylation
Term
Operator consitiuative mutants of the lac operon would ...
Definition
express beta-galactosidase constitutively
Term
E.coli two-component regulatory system
Definition
PhoB is a cytosolic protein
Term
How does the binding of the lac repressor to the lac operator block transcription initation
Definition
Lac repressor blocks RNA polymerase from interacting with DNA at the start site
Term
Which proteins "footprint" the lac operon control region?
Definition
Lac repressor, RNA polymerase, cAMP-CAP but NOT beta-galactosidase
Term
Fundamental difference between gene regulation in bacteria compared to eukaryotes
Definition
in eukaryotes trancription is influenced by how effectively the DNA sequence of promoter interacts with histone octomers
Term
CTD of RNA Pol II ... (3)
Definition
can be phosphorylated, is crutial for viability, and contains more than 50 repeats of heptapeptide

NOT present in RNA pol I II and III
Term
TATA box
Definition
acts to position RNA polymerase II for transcription initiation
Term
Eukaryotic promoters (3)
Definition
TATA box
initiator element
CpG islands
NOT an enhancer
Term
Correct order of binding transcription factor to RNA pol II promoters
Definition
TFIID, TFIIB, Pol II, and TFIIH
Term
What is the function of TFIIH?
Definition
unwinds the DNA duplex
Term
CpG islands
Definition
promoter for 60-70% of eukaryotic genes and a control region for genes that are transcribed at low rates
Term
Enhancer
Definition
DNA element that stimulates transcription of eukaryotic promoters
Term
elements used in EMSA?
Definition
radiolabled DNA fragment
polyacrimide gel
DNA binding protein
NOT DNase I
Term
Leucine Zipper motif contains
Definition
a leucine residue at every 7th position
Term
Strictual motifs of DNA binding domain
Definition
Homeodomain, zinc-finger, helix-loop-helix, NOT random coil acidic domain
Term
Heterochromatin (3)
Definition
stains more darkly with DNA dyes
contains more highly condensed DNA
associated with inactive genes

NOT susceptible to DNase I
Term
Mediator complex
Definition
has histone acetylase activity
Term
transcriptionally inactive genes
Definition
often are methylated
Term
lipid soluble hormones activate transciption by
Definition
binding to nuclear receptor
Term
Which protein domains are found in nuclear receptor family members
Definition
variable regions, DNA binding domain, and ligand-binding domain
Term
regulation of transcription by steroid hormones
Definition
involves cytoplasmic hormone receptors that can move to the nucleus
Term
X chromosome inactivation is mediated by
Definition
short RNA directed methylation of histones and DNA
Term
transcription initation and pol II activity (3)
Definition
does not require ATP, Pol II is responsible for synthesizing tRNAs and 5S-rRNA, and the promoter element of tRNA genes lie entirely within the transcribed sequence
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