2!!! - Control of gene expression: RNA Flashcards
What are isoforms?
Multiple proteins made from the same gene
What are 3 ways that isoforms made?
From:
- Alternative splice sites
- Alternative start sites
- Alternative polyadenylation (the end of the DNA)
What do some mRNAs have that can be regulated independently?
A second ORF
What 3 ways gene expression be regulated?
- Transcription
- Splicing
- Translation
- Regulated nuclear transport
- RNA stability
How many genes in the human are alternatively spliced?
75%
How many genes in the drosophila are alternatively spliced?
40%
What are 4 ways that alternative splicing can occur?
1) Optional exon
2) Optional intron
3) Mutually exclusive exons
4) Internal splice sites
What are mutually exclusive exons?
Exons that don’t occur at the same time
When do mutually exclusive exons usually occur?
When the exons code for similar proteins
What is an internal splice site?
Where the 3’ OR 5’ end splice site is in the middle of the intron instead of the end
How does the cell alternatively splice sites?
- Splice donor and acceptor sequences are very small (only 2 base pairs), meaning they are VERY FREQUENT
- Splice sites are relatively simple
- Mechanism isnt very precise - taken advantage of to produce different splice sites
How do RNA binding proteins assist alternative splicing?
They bind around the donor/acceptor splice sites and influence where splicing occurs
What affects the choice of splice site?
- RNA binding proteins
- Other sequences and secondary structure in the RNA
Why is the splicing mechanism not very precise?
To take advantage of it
What is the most common spliced gene?
Dscam in drosophila
What is Dscam involved in?
Specifying neurons in the brain of the drosophila
In what way is the Dscam gene alternatively spliced?
Has mutually exclusive exons
How many different possible isoforms are there in the Dscam gene?
38,000
What process is used to help to understand alternative splicing in humans?
Sex determination in drosophila
How is sex determined in drosophila?
By the amount of X chromosomes that the fly has:
- Male = X
- Female = XX
- This is then translated into sex characteristics
What are the three genes that regulate male or female differentiation?
1) sxl (sex lethal)
2) tra (transfomer)
3) dsx (double sex)
How are MALE characteristics determined in the drosophila?
- Transcripts sxl and tra are spliced to give INACTIVE protein isoforms
- dsx is ACTIVE
What does dsx do in the active form?
- Gives rise to a male specific repressor protein, which represses transcription from genes required for female development
How are FEMALE characteristics determined in the drosophila?
- MORE transcription of sxl due to two X chromosomes
- SMALL AMOUNT of functional sxl protein made using an alternative promoter
- sxl binds to its own transcript in a positive feedback loop to make MORE of the active protein by REPRESSING SPLICING at this site