Gene Expression I Test Two Flashcards

1
Q

How can an epidermal cell be different from a liver cell when they have the same genes?

A

They are different because they express their proteins differently

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2
Q

What are house keeping genes?

A

Genes that are present in all cells such as actin glucose, DNA replication transcription translation enzymes…

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3
Q

What two things does gene regulation require?

A

Short stretches of DNA sequences called recognition sites and Regulatory proteins

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4
Q

What is a LOGOS

A

demonstrates the flexibility of DNA binding proteins and what bases they can bind too. It is shown in a graph and the larger the letter represents the higher affinity.

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5
Q

Where are DNA recognition sequences located in regards to the first exon?

A

They can being either proximal or distal to the exon.

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6
Q

Where do most DNA binding proteins/motifs bind on DNA?

A

To the major groove (must be complementary). Usually make contact with the DNA in multiple places, the individual hydrogen bonds are weak but together they are strong.

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7
Q

What are the different domains found on a DNA motif?

A

Binding domain, dimer domain, activation domain, regulatory domain

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8
Q

Which two domains are not absolutely necessary?

A

Dimer and regulatory domains

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9
Q

Describe the experiment that proved a reporter and promoter gene exist? (Transcription Factors are modular).

A

DNA was put into a mouse and wild type DNA Beta galactose was produced. In the mutant type the first 50 AA were clipped off so no binding occurred. Another mutant strand had the C terminus cut away so even though the protein could bind there was no production of Beta Galactose (Activation).

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10
Q

What DNA Motif is connected by a short turn of amino acids to a short alpha helix and a long alpha helix, can be a dimer, and has a long alpha helix that recognizes the sequences on the major groove?

A

Helix Turn Helix

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11
Q

What is a Zinc Finger DNA Motif?

A

Zn atom in the center of two alpha helices, it is typically found in clusters, and each “finger” binds to the major groove of DNA.

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12
Q

Describe a Leucine Zipper.

A

2 Alpha helices interacting (Dimer, hetero or homo), similar to a clothespin when it binds DNA major groove, activation domain overlaps dimer domain, held together by hydrophobic interactions. between amino side chains (leu)

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13
Q

What DNA motif looks similar to a clothespin, has a short alpha helix connected with a loop of amino acids to another alpha helix. Also flexible, the helices can bend over one another and can be a hetero or homo dimer .

A

Helix Loop Helix

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14
Q

What is the most common and simplest DNA binding motif?

A

Helix turn helix

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15
Q

What separates dimers in a helix turn helix?

A

One turn

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16
Q

What domains are found in a Helix Loop Helix domain?

A

DNA binding, Dimerization and activation domains are found

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17
Q

Describe Hereditary Spherocytosis?

A

Mutation in the Zn finger Motif leading to a type of hemolytic anemia with round RBC’s. Caused from a mutation in a gene that codes for the EMS in RBC’s.

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18
Q

What is an example of a mutated zn finger protein that causes HS?

A

KLF1 zn finger protein binds to promoters of all genes in the EMS to turn them on. There are three exons involved, when a mutation occurs in this gene transcription does not occur and therefore not enough RNA is made so no protein is made.

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19
Q

What is the single nucleotide mutation in Hereditary Spherocytosis?

A

GAA to GAT changing Glu to Asp within exon three zn finger domain two. This will bind different and no transcription will occur.

20
Q

What makes up the Erythrocyte Membrane Skeleton?

A

Spectrin, anchorin, and alpha and beta subunits forming dimers and tetramers.

21
Q

What is EMSA? (Gel Mobility Shift Assay)

A

Electrophoresis on a protein gel using DNA mixed with protein. The DNA that binds to protein won’t travel far and can be found near the insertion site, while free DNA will move to the base.

22
Q

Affinity Chromatography?

A

Used to isolate DNA binding proteins. Purify the sequence specific binding proteins through salt baths. Start broad by having column with random DNA sequences add proteins. Then get specific using the particular sequence you are looking for.

23
Q

CHIP?

A

Chromatin Immuno-precipitation. Allows for hte identification of the sequence that a known protein binds too. Done in living cells and uses PCR to ID sequence at the end.

24
Q

What transcribes all protein coding genes?

A

RNA Polymerase II

25
Q

What is the promoter ?

A

Region where TF and RNA Pol II assemble.

26
Q

What are regulatory sequences?

A

Sequences that regulatory proteins bind to control the rate of assembly at the promoter.

27
Q

What allows the regualtory proteins to interact with proteins at the promoter?

A

DNA looping and the mediator

28
Q

How do gene activator proteins work?

A

They modify the DNA through changing the structure of chromatin and nucleosomes

29
Q

What do transcription activators bind to?

A

Bind DNA in nucleosomes

30
Q

What are the four ways that gene activator proteins modify DNA? How does this favor transcription?`

A
  1. Nucleosome Remodeling
  2. Nucleosome Removal
  3. Histone Replacement
    4.Histone modification
    Favors transcription because it makes DNA more accessible
31
Q

T or F: there are different sites for activators and repressors to bind to.

A

False- they compete for the same binding site on DNA

32
Q

How do gene repressor proteins inhibit transcription?`

A
  1. Blocks interactions with transcription factors.
  2. Recruits a chromatin remodeling factor to make the promoter return to a pre transcriptional nucleosome state.
  3. Repressors attracts a histone Deacetylase
  4. Repressor attracts methyl transferases
33
Q

Can proteins be both inhibitory and activator?

A

Yes. Depending on the make up of the complexes- regulation by committee

34
Q

How are gene regulatory proteins controlled?

A

Through synthesis, ligand binding, covalent modification, addition of subunit, unmasking and nuclear entry and proteolysis

35
Q

In hemoglobin what are the alpha globin like chains?

A

zeta and alpha

36
Q

In hemoglobin what are the beta globin like chains?

A

Epsilon
Gamma
Delta
Beta

37
Q

What are the fetal hemoglobin chains?

A

alpha and gamma

38
Q

What are embryonic hemoglobin chains?`

A

zeta and epsilon

39
Q

What are adult Hemoglobin chains?

A

Alpha and beta two of each

40
Q

How are the hemoglobin chains arranged?

A

In the order they are transcribed Z A A E G A D B

41
Q

How can Sickle Cell be cured?

A

CRISPR- repairing the single nucleotide change

Or if we could figure out how regulation of globin gene works we could turn back to fetal hemoglobin

42
Q

Differentiate two transcription factors.

Dr. White’s Question

A

Differentiate between Leucine Zipper, Helix loop helix, Zinc Finger, Helix turn Helix

43
Q

Describe one way TF are identified.

Dr. White’s question

A

Describe CHIP, Affinity Chromatography, EMSA.

EMSA: protein gel electrophoresis, DNA that binds to proteins will move slowly through the cell while free proteins move quickly to bottom allowing for sequencing to be determined.

44
Q

Describe two ways repressor proteins inhibit transcription.

Dr. Whites question

A

They can inhibit transcription by recruiting deacetylases or methy transferases to the area. They can also cause the nucleosomes to revert back to pre transcriptional form.

45
Q

Describe two ways in which gene regulatory proteins are controlled.

(Dr. White’s Question)

A

They can be controlled through covalent modification such as adding a phosphate. They can also be controlled through unmasking the inhibitor, stimulating nuclear entry by removing the inhibitory protein, adding another subunit…and more