Midterm 2 - Notes 1 (Part 1) Flashcards
Where can transposons be found? (2)
- Prokaryotes
2. Eukaryotes
What are 3 examples of transposons?
- Ac/Ds in maize
- P-element in drosophila
- IS element in E.coli
What does the protein coding region encode for?
Transposase
- site specific endonuclease DNA ligase
What 2 features do transposons have?
- Transposase
2. Two inverted repeated sections on either side of the transposase
What is an important hallmark for the inverted repeat sections?
That the repeats are short
What does length of the transposons depend on?
The target site
What do the protein coding regions contain?
Promoter
- drives the expression of the transposons
What does the transposase machinery do?
Cut out DNA/transposons and re-intergrate is somewhere else
What can you find inbewteen the two inverted regions?
Transposase
Transposase
Is an enzyme required for transposition of the IS element to a new site
- it catalyzes the process
IS
Insertion sequence
What does transposase do?
Ligates transposon into target site
What does cellular DNA polymerase and ligase do?
- Fills in overhangs
2. Joins ends
What does DNA polymerase and ligase create?
Short direct repeats specific for target sites
What happens when DNA polymerase and ligase are in the coding region?
It can be duplicated
- not shifted over
What does jumping (copy and paste) not lead to?
Increase in transposons copies
When can amplification only occur? If what 2 things happens?
During replication of host DNA
- Transposons jump from already replicated DNA to a region not yet replicated
- Transposons jumps in a germ line cell
Why do transposons need to be in a germ line? (2)
- So they can get passed onto generations
2. So they can increase in numbers
What happens if movement happens in the S phase?
Can have a chance of increase in numbers
What were the first transposons discovered?
Ac/Ds system affecting colour development in maize
What mechanisms do transposons act through?
Cut and paste mechanism
What does the 5’ end of a transposon do?
It is the recognition site which cuts them and creates a blunt end
What defines the frequency of a transposon
The expression level of a transposon gene
What creates sticky ends?
Transposase cuts randomly in the genomic DNA
What are 2 major classes of transposons?
- Transpose as DNA (DNA transposons)
2. Transpose vie RNA intermediate (retro-transposons)
What does transpose as DNA (DNA transposons) involve?
Tranposase
What mechanisms does transpose as DNA (DNA transposons) use?
Cut and paste mechanism
What does retro-transposons involve? (2)
- Reverse transcriptase
2. Integrase (similar to transposase)
What mechanism does retro-transposons use?
Copy and paste mechanism
How does the copy and paste mechanism work?
Make a copy of RNA and insert it somewhere else in the genome while also remaining in its original location
- just creates a copy of the original
What will transposons always do?
Amplify
What will the original transposon always do?
Stay where it was but duplicate
What are 2 major classes of retro-transposons?
- Retroviral-like retrotransposons
2. Non-retroviral-like retrotransposons
What do retroviral-like retrotransposons contain? (3)
- Long direct terminal repeats (LTR)
- at each end - Reverse transcriptase
- Integrase
LTR
Long Terminal Repeats
How do retroviral-like retrotransposons move?
Via an RNA intermediate whose production is driven by a promoter in the LTR
What are 4 examples of retroviral-like retrotransposons?
- Copia = drosophila
- Ty1 = yeast
- THE1 = human
- Bs1 = maize
What does non-retroviral-like retrotransposons contain? (4)
- AT-rich regions at flanking sites
- Non LTRs
- Reverse transcriptase
- Endonuclease
What is at the 3’ end of non-retroviral-like retrotransposon RNA transcript?
Poly A tail
What is at the 5’ end of non-retroviral-like retrotransposons?
Often truncated (shortened by cutting it)
How does non-retroviral-like retrotransposons move?
Via an RNA intermediate that is often synthesized from a neighbouring promoter
What are 3 examples of non-retroviral-like retrotransposons?
- F-element = drosophila
- L1 = human
- Cin4 = maize