Heterogenous Nuclear RNA
(hnRNA)
⚫ primary transcript
⚫ contains exons and introns
⚫ processed in the nucleus to mature
translatable mRNA or functional
tRNA and rRNA
The Anatomy of a Eukaryotic Gene
5' UTR
of mRNA coding sequence
Signal for site
of 3' end
trimming and Poly
(A) tail addition
Schematic drawing showing production of eukaryotic mRNA. The primary
transcript is capped before it is released. Then its 3′-OH end is modified, and
finally the intervening regions are excised. MeG=7-methylguanosine. **Two
nucleotides whose riboses may be methylated.
Post-transcriptional
Processing
Some Minor Processing Events
1. RNA Editing
– change in
the base sequence of an RNA
after it has been transcribed
e.g. mitochondrial mRNA in trypanosomes
-insertion of one or more U
http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/R/RNA_editing.gif
Some Minor Processing Events
2. Base Modification – the covalent
modification of a base within a
molecule
e.g. tRNA molecules
A few of the unusual nucleotides found
in tRNA molecules.
dimethyl G dihydro U isopentenyl A thiouridine
In most tRNA molecules about 10% of the nucleotides
are modified.
Processing of pre-mRNA
3 Major mRNA Processing
Events
1. Capping - addition of 5’ cap
2. Tailing - addition of 3’ polyA
tail
3. Splicing – removal of introns
1. Capping
addition of 7-methylguanosine triphosphate
to
the 5’ end
before transcription is over
capping is done by guanylyl transferase
stabilizes and prevents degradation of the
mRNA →enhances translation
ribosome recognition
Capping reaction
2. Tailing
polyadenylation - addition of up to 250
A’s to the 3’ end of the mRNA
Poly-A Polymerase (nuclear)
after removal of 3’end of the mRNA near
a conserved sequence ( AAUAA )
improves translation efficiency and
stability
Alternative Tailing
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
3. Splicing
▪ removal of segments (50-90%) from
primary transcript
▪ introns or intervening sequence are spliced out
▪ generally follows tailing
▪ caps retained, tails sometimes removed
Remember …
Prokaryotes: only tRNA
and rRNA are processed;
Polycistronic mRNA
transcripts
Eukaryotes: tRNA, rRNA
and mRNAs are
processed;
Monocistronic mRNA
mRNA Processing Example: in Euglena
mRNA Processing Example: human β-globin
Splicing Signals: Chambon’s Rule
▪ the 5' splice site → [ GU ] at the 5'
end of the intron
▪ the 3' splice site → [ AG ] at the 3'
end of the intron
▪ the branch point → located about 30
nucleotides from the 3' end,
consisting of just Adenine
Splicing Signals
introns begin with 5'-GU and end with 3'-AG
( GU - AG Rule )
mutant splice sites → aberrant splicing
Spliceosome
▪ large, nuclear RNA-protein complex
▪ catalyzes RNA splicing
▪ snRNAs complex with proteins to form
small ribonucleic particles ( “snurps” )
→ small nuclear ribonucleoprotein particles
▪ 5 snRNPs (U1, U2, U4, U5, and U6)
assemble on pre-mRNA to form
spliceosome
Role of snRNP’s in splicing
snRNP Size of Role
snRNA
(nucs)
U1 165 binds the 5’ splice site and then
the 3’ splice site
U2 185 binds the branch site and forms
part of the catalytic center
U5 116 binds the 5’ splice site
U4 145 masks the catalytic activity of U6
U6 106 catalyzes splicing
Two-Step Splicing-Reaction
Step 1: 1st transesterification reaction
→ cleavage of 5' exon-intron boundary
→ formation of intron lariat intermediate
Step 2: 2nd transesterification reaction
→ joins the two exons
→ release of introns
Alternative Splicing
▪ a way of introducing diversity to the
gene products of transcription
▪ creates proteins or RNA from
the same DNA sequence
RNA Transport
Eukaryotic
Prokaryotic
RNA Transport
eukaryotes
Nuclear pore complex
“more than just a hole”
energy requiring transport of molecules
into and out of the nucleus
Ran protein: energy generation
Karyopherins (exportins and importins):
transport
karyopherins
at least 20 different human karyopherins
each responsible for transport of a
different class of RNA
eg.
exportin t – for export of tRNAs in yeast
and mammals
importin B - for import of snRNAs from
cytoplasm into nucleus
Yra1p in yeast and Aly in animals – for export
of mRNA to their respective places in the cell
In bacteria, transcription is
simultaneous with translation.
“coupled transcription-translation”
Coupled
Transcription-Translation
end of transcription
