Post Transcription Flashcards
- primary transcript
- contains exons and introns
- processes in the nucleus to mature translatable RNA or functional rRNA and tRNA
heterogenous nuclear RNA (hnRNA)
where is hnRNA processed
nucleus
hnRNA is processed to mature into what
- translatable RNA
- functional rRNA and tRNA
AUG in eukaryotes is not necessarily found in __
+1
post-transcriptional processing
- minor processing
- major processing
minor processing events
- RNA editing
- base modification
change in the base sequence of an RNA after it has been transcribed
RNA editing
example of RNA editing
mitochondrial mRNA in trypanosomes
- insertion of one or more U
covalent modification of a base within a molecule
base modification
example of base modification
tRNA molecules
in most tRNA molecules about __ of the nucleotides are modified
10%
3 major mRNA processing events
- capping
- tailing
- splicing
addition of 5’ cap
capping
what is added in capping
7-methylguanosine triphosphate
addition of 3’ polyA tail
tailing
removal of introns
splicing
enzyme used in capping
guanylyl transferase
function of capping
- stabilizes and prevents degradation of the mRNA -> enhances translation
- ribosome recognition
how many As are added during tailing
up to 150 As
enzyme used in tailing
polyA polymerase (nuclear)
conserved sequence during tailing
AAUAAA
purpose of tailing
improves translation efficiency and stability
- some transcripts have 2 or more polyadenylation sites
- “choice” depends on factors such as developmental stage and location
- variability of proteins at post-transcriptional level is generated
alternative tailing
factors where alternative tailing depend on
- developmental stage
- location
- removal of segments (50-90%) from primary transcript
- introns/intervening sequence are spliced out
- generally follows tailing
- caps retained, tailes sometimes removed
splicing
percent removed during splicing
50-90%
spliced out during splicing
introns/intervening sequence
introns that are self-splicing
self-splicing group II introns (ribozymes)
what is processed in prokaryotes
- tRNA
- rRNA
what are processed in eukaryotes
- tRNA
- rRNA
- mRNA
prokaryotes do not have __
introns
- have many genes, producing multiple proteins
- only have one promoter
- have coregulation
- more efficient
polycistronic mRNA transcripts
- one protein: one sequence
- specific genes are turned on/off depending on cell needs, development, and environmental changes
- independent regulation
monocistronic mRNA
structure of introns after it is spliced
lariat
splicing signals
- 5’ splice site
- 3’ splice site
- branch point
5’ splice site
GU at 5’ end of intron
3’ splice site
AG at 3’ end of intron
Location of branch point
located about 30nt from 3’ end
what does the branch point consist
one adenine
introns begin with 5’-GU and end with 3’-AG
Chambon’s rule
mutant splice sites
aberrant splicing
- large, nuclear RNA-protein complex
- catalyzes RNA splicing
spliceosome
snRNA + protein
snRPS (small nuclear ribonucleoprotein particles)
5 snRNPS assemble to form what?
spliceosome
what are the 5 snRNPs
- U1
- U2
- U4
- U5
- U6
size:
U1
165
size:
U2
185
size:
U5
116
size:
U4
145
size:
U6
106
role:
U1
binds the 5’ splice site and then the 3’ splice site
role:
U2
binds the branch site and forms part of the catalytic center
role:
U5
binds the 5’ splice site
role:
U4
masks the catalytic activity of U6
role:
U6
catalyzes splicing
first splicesomes to attach to intron
- U1
- U2
two-step slicing reaction
- 1st transesterification reaction
- 2nd transesterification
what happens during 1st transesterification reaction
- cleavage of 5’ exon-intron boundary
- formation of intron lariat intermediate
what happens during 2nd transesterification reaction
- joins two exons
- release of introns
- a way of introducing diversity to the gene products of transcription
- creates proteins/RNA from the same DNA sequence
alternative splicing
example of alternative splicing
- calcitonin mRNA (thyroid)
- CGRP mRNA (hypothalamus)
- eukaryotes
- exits nuclear pore
RNA transport
energy generation
ran protein
exportins and importins
karyopherins
there are at least how many human karyopherins
20 different human karyopherins
example of karyopherins
- exportin-t
- Importin β
for export of tRNAs in yeast and mammals
exportin-t
for import of snRNAs from cytoplasm into nucleus
Importin β
for export of mRNA to their respective places in the cell of yeast
Yra1p
for export of mRNA to their respective places in the cell of animals
Aly
in bacteria, transcription is simultaneous with translation
coupled transcription-translation