EUKARYOTIC GENE CONTROL - Mr. Steckle's...
Transcript of EUKARYOTIC GENE CONTROL - Mr. Steckle's...
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EUKARYOTIC GENE CONTROL
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THE BIG QUESTIONS
• How are genes turned on and off? • How do cells with the same DNA/genes differentiate to perform completely different and specialized functions?
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GENE EXPRESSION
• The control of gene expression takes place along a specific pathway.
• 1. Packing/unpacking DNA • 2. Transcription • 3. mRNA processing • 4. mRNA transport • 5. Translation • 6. Protein processing • 7. Protein degradation/mRNA degradation
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1. DNA PACKING
• The double stranded DNA needs to coil and fold to fit inside of the nucleus.
• Double stranded DNA • DNA wraps around a histone protein
forming a nucleosome • Looped domains • Chromosome
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HISTONE/NUCLEOSOME
Positively charged amino acids on the histone proteins bind to the negatively charged DNA.
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LOOPED DOMAIN
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CHROMOSOME
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DNA PACKING CONTROLS TRANSCRIPTION
• Heterochromatin (H)-The DNA is tightly packed and will appear dark in an image.
• No transcription can take place so the genes in that region are “turned off”
• An example would be the insulin gene in
brain or bone cells.
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HETEROCHROMATIN
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DNA METHYLATION
• Methylation of DNA in the promoter region blocks transcription factors so the genes are “turned off”
• Occurs when a methyl group (CH3) is attached to a cytosine (C)
• This process in nearly permanent. • ex. Inactivation of mammalian X chromosome=Barr body
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DNA PACKING CONTROLS TRANSCRIPTION
• Euchromatin (E)-The DNA is loosely packed and will appear light in an image.
• Transcription can take place so the genes in that region are “turned on”
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EUCHROMATIN
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LOOSEN THE DNA BY HISTONE ACETYLATION
• Attachment of an acetyl group (COOCH3) to a lysine of the histone which will help unwind the DNA.
• The DNA will be loosely wrapped which will allow for transcription and the genes to be “turned on”.
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2. TRANSCRIPTION INITIATION
• Control regions of the DNA • Enhancer is a short region of DNA bound by
regulatory proteins to increase the likelihood of transcription.
• Promoter is a nearby control sequence of DNA where transcription factors and RNA Polymerase can bind. This is called the transcription complex.
• The DNA will fold forming the transcription complex and the mRNA can begin to be produced.
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3. POST TRANSCRIPTIONAL CONTROL
• Removal of introns using spliceosomes.
• Add a 5’ G cap
and 3’ poly A tail.
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4. REGULATION OF MRNA DEGREGATION
• Life span of mRNA determines the amount of protein synthesis.
• The mRNA can last from hours to weeks.
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DEATH OF MRNA
• Small interfering RNA (siRNA)
• Short segments of RNA that bind to mRNA and create double stranded mRNA.
• Triggers the cell to degrade the mRNA. The “death tag” for mRNA.
• Causes gene silencing. The gene is turned off so no protein is produced.
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ACTION OF SIRNA
siRNA
double-stranded miRNA + siRNA
mRNA degraded functionally turns gene off
Hot…Hot new topic in biology
mRNA for translation
breakdown enzyme (RISC)
dicer enzyme
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5. CONTROL OF TRANSLATION
• Block initiation of translation tag • Regulatory proteins attach to the 5’ end of mRNA • Prevents the attachment of ribosomal subunits
and initiator tRNA • Blocks translation of mRNA to protein
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6. PROTEIN PROCESSING AND DEGRADATION
• Protein processing • Folding, cleaving, adding sugar groups and
targeting for transport.
• Protein degradation • Ubiquitin-Tags the protein for degradation by
proteases • Proteasomes-Responsible for the degradation of
the protein.
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UBIQUITIN
• “Death Tag” • A 76 amino acid polypeptide that
marks unwanted proteins with a label to be broken down by proteasomes.
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PROTEASOMES
• Protein-degrading machine. • Breaks down any protein into 7-9
amino acid fragments that can be recycled.
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THE WHOLE STORY
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initiation of transcription
1
mRNA splicing
2
mRNA protection 3
initiation of translation
6
mRNA processing
5
1 & 2. transcription - DNA packing - transcription factors 3 & 4. post-transcription - mRNA processing - splicing - 5’ cap & poly-A tail - breakdown by siRNA
5. translation - block start of translation
6 & 7. post-translation - protein processing - protein degradation
7 protein processing & degradation
4
4
Gene Regulation