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MCB Transcription/RNA process
by Chelsea (complete)
30
Medical
Graduate
02/20/2009

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Term
Why is RNA less stable than DNA?
Definition
The 2' hydroxyl group can act as a nucleophile to attack the phosphate (a "juicy target") of the phosphodiester backbone, causing hydrolysis. Also, RNA can't make a B-form when in helical structure, and so it makes the more squished A-form.
Term
Typically, what types of RNA are long-lived?
Definition
rRNA and tRNA. mRNAs are usually meant to "destroy after reading" (very James Bond style...)
Term
What recently discovered types of RNA are used to regulate stability or expression of mRNAs?
Definition
Micro RNAs (miRNAs) and short interfering RNAs (siRNAs)
Term
Imagine the DNA helix. Which strand (the coding or template strand) will have the same sequence as the RNA that is being made?
Definition
the Coding strand! (which is displaced). NOT the template strand.
Term
What is the Pribnow box?
Definition
Where Prokaryotic RNA polymerase jumps on the DNA promoter region and makes a stable, inert closed complex.
Term
How is rifampicin used?
Definition
Only Prokaryotic RNA Polymerase it sensitive to it, so it can be used as an antibiotic.
Term
What region of the histones (H3 and H4) are acetylated or methylated?
Definition
The "histone tails" of the N-terminal tails of histones. These areas don't contribute to the formation of the nucleosome core, and don't alter stability of the histone-DNA interactions.
Term
What 4 enzymes add or remove acetyl and methyl groups from the histones?
Definition
HATs, HDACs, HMTases, histone demethylases.
Term
Zolinza (vorinostat) is an HDAC inhibitor that is FDA approved for treatment of cutaneous T-cell lymphoma. How does it work?
Definition
Inhibiting HDAC results in hyperacetylation, and is somehow worse for tumor cells than it is for normal cells. Maybe the result is hyperacetylation of tumor suppressor gene that reduces cell growth more specifically in cancer cells.
Term
What do bromodomains do?
Definition
They recognize acetylation on the H teminal H4 tail and "bring friends" (the remodeling complex).
Term
What is the "histone code"?
Definition
It is the pattern of modifications. How many Acs do you have? How many Mes? How many Ps? These combinations interact to mean different things.
Term
What happens if you have a mutation in the MeCP2 gene?
Definition
Leads to Rett's Syndrome. Normally, MeCP2 binds to methylated (inactive) DNA and calls in HDAC (which un-acetylates and inactivates even more!). If you don't have MeCP2, you don't silence areas that are supposed to be totally inactive.
Term
Decitabine completed a Phase I trial for inhibiting growth of certain tumor types. How does it work?
Definition
When tumor suppressor genes are methylated, they are silenced and growth goes crazy! Decitabine looks like a cytosine, but doesn't have a carbon at the 5' position (it has a nitrogen instead). This nitrogen can't get methylated!
Term
What is the difference between "cis" and "trans" elements?
Definition
"Cis" elements (like promoters) only affect the expression of the DNA they are embedded in. "Trans" elements (like binding proteins) are encoded in one place, but affect any DNA molecule in the cell that has their recognition site.
Term
RNA polymerase cannot recognize a Eukaryotic promoter sequence. How does it know where to go?
Definition
It recognizes the combination of DNA binding proteins, general transcription factors, TBP, mediator. (Order differs from promoter to promoter)
Term
What do General Transcription Factors (GTFs) do?
Definition
They make a landing zone for the RNA polymerase. If they don't create a "suitable platform," RNA Polymerase will keep cruising around. They do this by calling in TBP to the TATA box and mediator to the promoter.
Term
Prokaryotic promoters are all very similar. What does this tell us about one promotor?
Definition
It can control multiple genes in an operon. Also, in prokaryktes, the strength of a promoter is mostly controlled only by how well it matches the sigma factor binding site.
Term
What does it mean to act "in cis"?
Definition
A change in the promoter sequence will affect how often the gene on the DNA molecule that is altered is transcribed. How most promoter mutation act.
Term
What does it mean to act "in trans"?
Definition
The factor will affect aLL DNA molecules in the cell (not just the one that encodes it). How most DNA binding proteins that recognize promoter elements act.
Term
In prokaryotes, what do operators do?
Definition
They modulate promoter activity, and are usually very close to the promoter (or can overlap).
Term
p53 is usually very unstable and gets degraded soon after it is made. If DNA in a cell is damaged, what happens to the signal to degrade p53?
Definition
It gets interrupted. The protein builds up, and gets transported to the nucleus where is acts as a transcriptional activator. bad news! cancer!
Term
In eukaryotes, transcription factors are often regulated how?
Definition
By altering their stability or subcellular location.
Term
Why do we use Northern Blots?
Definition
To look at size/structure/abundance/tissue distribution of mRNA.
Term
Why do we use a microarray?
Definition
To detect chromosomal changes. A microarray can see changes in gene expression patterns. It uses LOTS of messages at the same time.
Term
What does Drosha do? How does this relate to Dicer?
Definition
It is an endonuclease that digests dsRNA hairpins in the nucleus. These small fragments are transported to the cytoplasm where they are digested even more by Dicer. End product is miRNAs.
Term
How does RISC work?
Definition
It uses the miRNA as a guide to stop translation of mRNAs with the same sequence.
Term
What must be on the 3' end of all tRNAs?
Definition
CCA
Term
What does poly-A polymerase do?
Definition
It is an RNA polymerase that adds a poly-A tail to the 3' end of mRNAs. It works without a template.
Term
What are ADARs?
Definition
Convert specific adenosines to inosines. Inosine pairs with cytosine. This is an example of how bases within the RNA can be modified.
Term
ApoB48 is shorter than ApoB-100 due to a point mutation. What causes this?
Definition
Cytosine deamination. The mRNA for ApoB-48 is processed to change the CAA codon to UAA, resulting in premature termination of translation.
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