Gebelein 1 Flashcards

Flashcards for Gebelein Lecture 1

1
Q

_____ are a gene profiling technique where cDNA is hybridized to a plate with complementary oligos to assess gene expression levels.

A

Microarrays

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2
Q

A classic gene profiling technique, the _____ blot, uses a labeled probe to detect a specific transcript.

A

Northern

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3
Q

_____ is a gene profiling technique where mRNA is converted to cDNA and amplified with PCR to measure gene expression.

A

qPCR

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4
Q

_____ is a powerful gene profiling technique that involves isolating mRNA, converting it to cDNA, and sequencing it.

A

RNA sequencing

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5
Q

Single-cell RNA sequencing allows researchers to analyze gene expression at the _____ level.

A

individual cell

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6
Q

One advantage of RNA-seq over microarrays is that it allows for the analysis of sequence _____, such as mutations.

A

changes

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7
Q

Eukaryotic gene expression can be controlled at _____ steps.

A

six

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8
Q

RNA _____, which encompasses splicing, localization, and degradation, represents a key control point in gene expression.

A

processing

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9
Q

_____ are DNA sequences that can recruit sequence-specific transcription factors to enhance or inhibit the activity of RNA polymerase at the promoter.

A

Enhancers

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10
Q

_____ are DNA sequences responsible for recruiting the RNA polymerase machinery and initiating transcription.

A

Promoters

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11
Q

_____ function as boundaries on a chromosome, preventing interactions between DNA sequences located on opposite sides.

A

Insulators

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12
Q

Proteins that bind to specific DNA sequences (TFBSs) to regulate transcription are known as _____.

A

Sequence-specific TFs

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13
Q

Proteins that bind to specific DNA sequences (TFBSs) to regulate transcription are known as _____.

A

Sequence-specific TFs

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14
Q

General TFs, together with RNA polymerase II, are core proteins essential for the synthesis of all _______.

A

mRNAs

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15
Q

_____ are proteins that package DNA into structural units called nucleosomes.

A

Histones

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16
Q

Modifications to histones can alter their ability to bind to _____.

A

DNA

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17
Q

Regulatory proteins known as _____ are recruited by transcription factors to either activate or repress transcription, often through modifications to histones or the RNA pol II enzyme.

A

co-activators/co-repressors

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18
Q

_____ assays are one method used by researchers to assess for promoter activity in cells.

A

Reporter

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19
Q

_____ and _____ are two approaches used to determine which transcription factor binds to a particular DNA sequence.

A

Biochemical, bioinformatic

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20
Q

The initial crucial step in transcriptional regulation is the recognition and ________ of a pre-initiation complex (PIC) at promoter sequences.

A

recruitment

21
Q

The second step in transcriptional regulation, _____, encompasses the “melting” of DNA to form an open complex and the phosphorylation of Ser-5 on the CTD of RNA polymerase II.

A

transcription initiation

22
Q

_____ involves the phosphorylation of the CTD at both Ser2 and Ser5.

A

Transcription elongation

23
Q

Key components involved in promoter recognition and PIC formation include: _____, general transcription factors (TFIID, TFIIA, TFIIB, TFIIE, TFIIF, TFIIH), and core promoter sequences.

A

RNA polymerase II (RNA-polII)

24
Q

Scientists utilized _____ genetics and biochemical fractionation coupled with protein complex _____ to initially identify and investigate the key “trans” players in transcription.

A

yeast, purification

25
Q

While the biochemical definition of the PIC excludes the _____ complex, it is important to note that the _______ is essential for general transcription in vivo.

A

mediator

26
Q

The composition of some ________ TFs, such as TFIID and the Mediator, can vary among cell types due to the expression of distinct subunits.

A

“general”

27
Q

_____ is a technique used to identify “protected” (bound) DNA sequences by detecting regions where protein binding shields DNA from nuclease digestion.

A

DNA footprinting

28
Q

_____ is a technique employed to study the interactions between proteins and DNA within a cellular context.

A

Chromatin Immunoprecipitation (ChIP)

29
Q

Compared to footprinting with a nuclease, ChIP offers _____ resolution.

A

lower

30
Q

Two variations of the ChIP technique are _____ and _____.

A

ChIP-PCR, ChIP-seq

31
Q

_____ combines ChIP with exonuclease digestion, enabling the identification of protected DNA regions in vivo with enhanced precision.

A

ChIP-Exo

32
Q

A crucial DNA-binding component of the TFIID complex, the _____ (TBP), plays a vital role in transcription initiation.

A

TATA Binding Protein

33
Q

TBP binds to the _____ groove of DNA, resulting in both a local distortion of the DNA path and partial unwinding of the DNA helix.

A

minor

34
Q

_____ possesses both helicase activity, essential for unwinding the DNA double helix to create the transcription bubble during initiation, and kinase activity, responsible for phosphorylating the CTD of Pol II.

A

TFIIH

35
Q

The _____ complex plays a crucial role in recruiting the super elongation complex (SEC) after the release of PolII from the PIC, marking a key step in the transition from transcription initiation to elongation.

A

Mediator

36
Q

What is gene regulation?

A

The “controlled” making of gene products required for cellular function.

37
Q

Mammalian DNA contains around ________ genes.

A

20,000

38
Q

What are the major control points in gene regulation?

A

Transcription, RNA processing, Translation, Post-translational modifications and degradation

39
Q

What are cis-regulatory modules?

A

DNA sequences that are required for and/or alter the process of transcription.

40
Q

What are promoters?

A

DNA sequences that recruit the RNA polymerase machinery and initiate transcription

41
Q

What are enhancers?

A

DNA sequences that recruit sequence-specific transcription factors that ”enhance” or ”inhibit” the activity of RNA polymerase at the promoter.

42
Q

True or False: Enhancers must be near promoters.

A

False, they can be distant from promoters

43
Q

What are insulators?

A

DNA sequences that recruit “architectural” proteins to create barriers along the chromosome. In general, DNA sequences on opposite sides of the insulator
do not physically associate with each other.

44
Q

Areas between insulator sequences are often called ___________.

A

Topological associated domains (TADs)

45
Q

What are transcription factor proteins?

A

Proteins that regulate the
process of transcription

46
Q

What are the core proteins required for all mRNA synthesis?

A

General” TFs and RNA polII

47
Q

What are sequence-specific TFs?

A

Proteins that bind specific DNA
binding sites (TFBSs) to aid or inhibit transcription. Can
bind either promoters or enhancers.

48
Q

What are co-activators/co-repressors?

A

Regulatory proteins
recruited by TFs to either promote or inhibit transcription,
often by modifying histones or the RNA polII enzyme.

49
Q
A