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Genetics > Regulation of Gene Expression in Eukaryotes > Flashcards

Regulation of Gene Expression in Eukaryotes Flashcards

1
Q
  1. Change how much mRNA is produced
  2. Change mRNA sequence
  3. Get rid of mRNAs that have already been made
  4. Change how much protein is produced from mRNA
  5. Modify protein function
  6. Degrade proteins that have already been made
A

Strategies used by eukaryotes to regulate gene expression

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2
Q

region of mRNA upstream of start codon that is transcribed but not translated

A

5’-UTR

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3
Q

region of mRNA downstream of stop codon that is transcribed but not translated

A

3’-UTR

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4
Q

required for basal transcriptional activity; it is always adjacent and upstream to the coding sequence it controls and its function is directional

A

Promoter

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5
Q

Influence rate of transcription, bound by transcription factors (activators or repressors)

Can regulate transcription at a distance, can be upstream, downstream, or within a gene, and are orientation-independent

A

enhancers/silencers

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6
Q

Forms a “bridge” between transcriptional activators/repressors and RNA polymerase

A

mediator

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7
Q

Proteins that help bridge between transcription factors and GTFs

A

coactivators

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8
Q

Domain: Recognizes specific DNA sequence

A

DNA-binding domain

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9
Q

Domain: recognizes and binds to other proteins

A

dimerization domain

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10
Q

Domain: binds to small molecule to activate transcription factor activity

A

ligand-binding domain

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11
Q

recognizes and binds to other transcriptional machinery

A

activation/repression domain

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12
Q
  • used to test the expression of a small number of genes of interest
  • principle is the same as PCR, but with a cDNA library (DNA made using RNA template) as a template and a fluorescence-based mechanism to measure amplification over time
  • Sybr-green: dye that fluoresces only when incorporated into dsDNA
    -Amount of RNA for a gene of interest is determined by measuring how many cycles (each cycle doubles material) it takes to reach a given fluorescence intensity
  • the fewer cycles, the more starting material
A

Measuring levels of gene expression: Quantitative reverse transcriptase PCR (qRT-PCR)

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13
Q
  • total RNA is isolated from cells, converted into cDNA library, and sequenced
    -sequencing methods generate large number of short sequencing reads that correspond to cDNA
  • the number of sequencing reads generated for a particular gene reflect the quantity of RNA produced for that sequence in the original sample
A

RNA-sequencing (RNA-seq)

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14
Q

-used for visual information about gene expression
-regulatory region of a gene is fused to the coding sequence of a protein that, when expressed, produces a visual cue

  • used to identify regulatory sequences
  • fluorescence intensity or enzyme activity can be used to quantify gene expression
A

reporter transgenes

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15
Q

DNA is wrapped around histone octamers

A

nucleosomes

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15
Q

-Electrophoretic Mobility Shift Assay (EMSA) “Gel Shift”

  • Chromatin Immunoprecipitation (ChIP)
A

Used to identify transcription factor binding sites

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16
Q

Made up of two each: histone H2A, histone H2B, histone H3, and histone H4

A

histone octamers

17
Q

results in less compact chromatin structure and is associated with transcriptionally active DNA

A

Acetylation

18
Q

can be recruited by activators and acetylates histones

A

Histone Acetyl Transferase (HAT)

19
Q

can be recruited by repressors and removes acetyl groups from histones

A

Histone Deacetylase (HDAC)

20
Q
  • Eukaryotes methylate cytosines to silence gene expression
    -Eukaryotic DNA methylation occurs at ___
A

CpG islands

21
Q

Silences gene expression, occurs between adjacent C and G bases, not a single C/G base pair

A

Eukaryotic Methylation

22
Q

Catalyzes DNA methylation,
can methylate DNA de novo, or add methyl groups to daughter DNA strands during DNA replication when parental molecules are methylated (maintenance)

A

DNA methyltransferases (DNMTs)

23
Q

Alter chromatin structure:
- sliding
- ejection or replacement of histone octamer
- ejection or replacement of an H2A-H2B dimer
exchange of a variant histone

A

Reader Proteins

24
Q

heritable changes in gene expression not attributable to nucleotide sequence variation

A

epigenetics

25
Q

inheritance of gene expression between generations

A

genomic imprinting

26
Q

regions of silenced DNA

A

imprints

27
Q
  1. Imprints are inherited from parental cells
  2. imprints are erased during meiosis
  3. New imprints are made based on gametogenesis
A

Steps of genomic imprinting

28
Q

Alternative splicing produces different proteins

A

isoforms

29
Q

removal of 5’ cap

A

decapping

30
Q

removal of 3’ polyA tail

A

deadenylation

31
Q

Targets mRNAs for degradation

A

siRNA

32
Q

cytoplasmic protein that recognizes double-stranded sRNA or pre-miRNAs and cuts them into ~21-24nt segments

A

Dicer

33
Q

contains the AGO (Argonaute) protein and serves as a programmable search engine; when it finds a guide strand match it cleaves the mRNA

A

(RISC) RNA-induced silencing complex

34
Q

control mRNA stability and translation

a single ____ can inhibit the translation of many mRNAs

A

microRNAs (miRNAs)

35
Q

Phosphorylates proteins

A

kinases

36
Q

dephosphorylates proteins

A

phosphatases

37
Q

results in conformational change of protein and impacts its function (can activate, repress, or alter protein activity)

A

Phosphorylation

38
Q
  • major function is as a protease

Targets:
- misfolded proteins
- proteins that are regulated
- proteins no longer needed
- cleaved protein fragments

A

proteasome

39
Q

proteins targeted for degradation are _________ by regulatory enzymes called ubiquitin ligases

A

ubiquitinated

Genetics (9 decks)

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