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Flashcards in 8.16.16 Lecture Deck (49)
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1

Gene expression must be ___. This is the basis for what three things?

Regulated; Development, homeostasis, and pathogenesis

2

True or false - all cells have the same genome.

True

3

Different types of cells express different sets of ___ that determine the ___ and ___ of the cell type.

Genes; character; function

4

What is the transcriptome?

All mRNAs expressed by a cell

5

What is the proteome?

All proteins expressed by a cell

6

Genes can be regulated with respect to what three things?

Time, space, and quantity

7

What is the promoter region and what does it do?

Made up of the core promoter (-40 to +40) and the proximal promoter (-200 to +50); promotes transcription in an orientation and distance-dependent manner

8

What does the core promoter do?

Induces basal expression of a gene; the location of assembly of general TF and polymerase

9

What are regulatory elements and what do they do?

Enhancers and silencers; short DNA sequences that bind transcription factors/regulatory proteins that interact with the basal transcription machinery in a distance and orientation independent manner when outside of the proximal promoter region (can be upstream or downstream of start site, or within introns/exons). Note that when they are within the proximal promoter region, they are dependent on distance and orientation.

10

What affects the rate of initiation?

Regulatory elements

11

What are insulators and what do they do?

Short DNA sequences found at the ends of a gene unit, bind insulator proteins, contain signals in a unit, block influence of outside regulators, prevent heterochromatin migration

12

What allows regulators bound anywhere on a gene to interact with proteins assembled at a promoter? Many regulators act through ___. Others assemble directly at the promoter.

DNA looping; Mediator

13

True or false - each regulatory element may only bind one transcription factor.

False - multiple TF may be bound to one regulatory element

14

Alterations in chromatin structure are directed by eukaryotic ___.

Transcription activator proteins

15

Promoters are initially buried in ___.

Heterochromatin

16

At least one ___ is bound on the surface of heterochromatin to begin the process of transcription.

Transcription regulator

17

What are three possible alterations to chromatin structure that promote gene transcription by increasing the accessibility of DNA and facilitating binding of RNA polymerase and general transcription factors?

1. ATP-dependent chromatin remodeling complex causes nucleosome sliding to open up access to DNA
2. Histone chaperones - removing histones, creating nucleosome-free DNA, or replacing histones with variants to favor euchromatin
3. Histone-modifying enzyme - specific modification patterns destabilize heterochromatin and attract components of transcription.

18

What is one potential order of gene activation? Note that this differs from gene to gene.

1. TF binds to chromatin
2. Chromatin remodeling occurs via the remodeling complex
3. Covalent histone modification occurs via histone modification enzymes
4. Additional activator proteins bind to the gene regulatory region
5. Assembly of pre-initiation complex at the promoter (Mediator, general TF, RNA polymerase)
6. Transcription initiation (other gene activator proteins, rearrangement of proteins in pre-initiation complex)

19

Describe acetylation and methylation of Lysine.

-Acetylation removes the positive charge of Lysine, which breaks its interaction with negatively charged DNA.
-Methylation creates binding sites for proteins.

20

How do histone acetylation and deacetylation occur?

Histone acetyl transferase (HAT) catalyzes acetylation, generating euchromatin. Histone deacetylase (HDAC) catalyze deacetylation, generating heterochromatin.

21

What does histone methylase do?

Add CH3 to Lys and Arg

22

What does histone kinase do?

Phosphorylate Ser

23

Each histone modification attracts ___ that specifically bind to the modified site. What are the two major types?

Proteins; Bromodomain (binds to acetylated Lys and Arg, and phosphorylated Ser) and Chromodomain (binds to methylated Lys and Arg)

24

Give an example of successive histone modifications during transcription initiation. Note that these steps vary in various genes.

1. Transcription activator protein and HAT bind to DNA.
2. HAT acetylates Lys.
3. Histone kinase binds and phosphorylates Ser.
4. Further acetylation
5. TFIID and chromatin remodeling complex bind chromatin (recognize acetylated histone tails through bromodomain)
6. Remainder of transcription machinery assembles; transcription begins.
*Note that coactivation occurs throughout

25

What do transcription factors do?

Induce transcription

26

What are the properties of transcription factors?

1. Modular designs with three domains
2. DNA binding domain contains a structural motif with amino acids that interact with a unique DNA element exposed by the major and/or minor groove of the helix.
3. Commonly homo- or hetero-dimers
4. Dimers bind to a palindrome in the DNA, which contain a symmetry appropriate for dimer binding to two successive turns in the DNA helix.

27

What are the three domains typically found in transcription factors?

1. Activation (or repressor) domain
2. Dimerization domain
3. DNA binding domain

28

TF generally interact with the ___ groove of DNA. Why?

Major; it has more exposed information (more available bases)

29

What are the four common motifs found in DNA binding proteins?

1. Helix-turn-helix (HTH)
2. Helix-loop-helix (HLH)
3. Zinc finger (Zn finger)
4. Basic Leucine zipper (b-ZIP)

30

What is the recognition helix of HTH?

C-terminal alpha-helix; participates in sequence-specific recognition of DNA (contains DNA binding domain); fits into the major groove.