Study BMB 400 Chapter 11 Flash Cards

 
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BMB 400 Chapter 11

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LINE:
Long inter-spread nuclear element, autonomous poly-A retrotransposons, ~20% human genome, L1, integrase and reverse transcriptase
-represent reverse-transcribed RNA molecules originally transcribed by RNA polymerase II into mRNA
SINE:
Short inter-spread nuclear element
-nonautonomous
-poly-A retrotransposons, Alu, no integrase and no reverse transcriptase
-LINE-encoded proteins bind immediately to their own RNA during translation
-represent reverse-transcribed RNA molecules originally transcribed by RNA polymerase III into tRNA, rRNA, and other small nuclear RNAs.
Yeast Ty Elements
Transpose into Safe Havens in the Genome
-Transposons in Yeast
- viral-like retrotransposons
-preferentially integrate into specific chromosomal regions
Mechanism: formation of specific protein-protein complexes between the integrase and host specific proteins bound to these chromosomal sites (Sir4 for Ty5). Try to integrate a site away from important genes for host. This is specially important with small, gene-rich genomes (e.g., yeast)
Tcl/Mariner Elements
-DNA elements in Eukaryotes
-simplest autonomous transposons -only a pair of terminal inverted repeat sequences and a gene encoding a transposase protein
-move by a cut-and-paste transposition mechanism
Artificially hyperactivated
-Sleeping beauty
-promising as a tool for mutagenesis and DNA insertion in many eukaryotic organisms
MuB:
ATPase that stimulates MuA activity and controls the choice of the DNA target site
MuA:
transposase, member of the DDE protein superfamily
Tn10 Transposition
-Coupled to Cellular DNA Replication
DNA methylation regulates Tn10 transposition
Limit the overall frequency of transposition
It also restricts transposition specifically to actively dividing cells
Two copies of the chromosome present to heal the double-stranded DNA break left in the old target site
IS4-Family Transposons
Are Compact Elements with Multiple Mechanisms for Copy
Number Control
Antisense RNA controls the expression of the transposase gene to regulate copy number
Poly-A Retrotransposons
Example: human LINE elements.
The poly-A retrotransposons also move by using an RNA intermediate
Many of the poly-A retrotransposons that have been detected by genomic sequencing.
Most of these elements are truncated therefore have lost the ability to transpose.
Viral-like Retrotransposons and Retroviruses
-transposon transcribed into mRNA
-reverse transcriptase
-interase-catalyzed 3' end cleavage
-this RNA does not carry the entire LTR sequence
Replicative transposition .
frequently causes chromosomal inversions and deletion (disadvantage for a selection)
DNA transposition
-cut and paste
-replicative
There Are Multiple Mechanisms for Cleaving the Nontransferred Strand during DNA Transposition (
-transposons cleave 3' end of eement DNA
-different ways to cleave 5' end
cut and paste
Transposase recognizes terminal inverted repeats at the end of transposon and brings the two ends together to generate stable protein-DNA complex (synaptic complex or transpososome)
Elements of DNA transposons
terminal inverted repeat (recombinase sites)
transposase (integrase)
additional genes like AB resistance
transposon frequency
More than 50 % of both the human and maize genomes are composed of transposon-related DNA sequence
Ftks
-cell division protein
-if FTKs unavailable, only 2 XerC active and cleave
-can't fully separate
-goes back to original position a lot
-when FTKs is available, 2nd pair of XerD cleaves and separates
Xer
-tyrosine kinase
-cell division protein: Ftsk
-XerC and XerD
-XerC always cleaves
-need FTKs for XerD to work
Resolvases (site-specific recombinases)
generate monomeric DNA molecule from dimer.
How resolvases carry out directional selectivity (coupled with cell division) of the recombination reaction?
Site-specific DNA bending protein, Fis
stimulates the rate of recombination ~1,000-fold (When Fis protein is present, the three-segment complex “invertasome” can form, Fig. 11-14).
Salmonella Hin recombinase
is common in bacteria, known as programmed rearrangement. It changes expression of two alternative form of flagellin (H1 and H2 forms).
Phage l Excision Requires a New DNA-Bending Protein
(called architectural protein)
Tyrosine Recombinases
requires four subunits of Cre, each subunit bound to one binding site on the substrate DNA molecules
-Two fold symmetry instead of four (“one strand at a time” exchange mechanism).
-active sites switch thing
Tyrosine Recombinases
-makes 2 breaks
-Cleave and rejoin two DNA strands first and then cleave and region the other two strands
Four molecules of recombinase are needed
-Holiday junction intermediate
Serine Recombinases
-DSBs in both DNAs
-cleave all four strands
A minimum of four subunits (that is a tetramer) of the recombinase is required
-just flip the DNA after cleavage
serine recombinase; and 2) tyrosine recombinases.
-covalent protein-DNA intermediate
-site specific
-no external energy (e.g. ATP-hydrolysis) is needed for DNA cleavage and rejoining.
Site-Specific Recombination
insertion, delietion, inversion
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