Sie sind auf Seite 1von 21

Week of MAR-7

Spring Break
(next week)

1
221.S16
Week of MAR-14

M T W R F
10-11 10-11
Iryna Run
office hours office hours
LLAB 114 LLAB 114
11:00-11:50 noon-1:00 11:00-11:50 11:00-11:50
Class GG Class Class
LLAB 10 office hours LLAB 10 LLAB 10
Questions? LL114
LLAB 319
12-1
Steve
office hours
LLAB 114
5-6:30 6:30-8:00 6:30-8:30 5:15-6:15
Review Review Exam-2 GG
LLAB 10 LLAB 10 LLAB 10 office hours
LLAB 319

2
221.S16
What is on Exam-2?
Content
• The primary emphasis for the exam will be the material that we covered in
class from Monday 2/8/16 (Allele interactions) through Wednesday 3/2/16
(RNA splicing).

• You are also responsible for the corresponding reading in the text (see
syllabus) and for the required reading listed on our Canvas site.

• You should also have a good understanding of the material & problems
corresponding to the prerequisites for the course (see syllabus).

• As you might imagine, you should be pretty facile at solving problems like
those included on the syllabus and in the recitations.

Review sessions
• Monday 3/14 5:00-6:30 LLAB 10
• Tuesday 3/15 6:30-8:00 LLAB 10

Exam-1
• Wednesday 3/16 6:30-8:30 LLAB 10
• Simple calculators are permitted for this exam. Laptops, iPads, PDAs,
3
smartphones, etc. are NOT permitted
221.S16
Clarification

Shown below is a 50-nucleotide double-stranded DNA segment from a prokaryotic genome that contains a
factor-independent transcriptional terminator element.

5’-...GCTTTACAAAAAAAGTGCGCTGCTGCTTAGCAGCGCACACCTTGGTGGGG...
3’-...CGAAATGTTTTTTTCACGCGACGACGAATCGTCGCGTGTGGAACCACCCC...

a. Write the nucleotide sequence of the transcribed RNA. There is no need to indicate any secondary
structure but clearly label the 5’- and 3’-ends of this RNA.

b. If the blue-shaded 3-bp sequence were inverted, would this affect the efficiency of transcriptional
termination? Briefly explain.

3’-CGA-5’ This is what I had in mind for an “inversion.” Basically,


5’-GCT-3’ rotating the sequence so the sequence so the strand
polarities are maintained.

c. If the red-shaded 2-bp sequence were deleted, would this affect the efficiency of transcriptional
termination? Briefly explain.

4
221.S16
Anatomy of an mRNA transcript

5’-untranslated region (leader) 3’-untranslated region (trailer)

H2N
COOH

[ORF: open reading frame]

5
[UTR: untranslated region]
221.S16
Translational initiation: Prokaryotes

ribosome binding site (RBS)

3’

5’

How does the ribosome find the AUG? In prokaryotes, a conserved


sequence (Shine-Dalgarno) near the 5’ end of the mRNA binds to a
complementary sequence in the 16S rRNA of the 30S (small) ribosomal
subunit, aligning the AUG codon in the “P” site to start the chain.
6
221.S16 B. Keith
Translational initiation: Prokaryotes

Once the AUG is properly aligned,


bacterial protein synthesis begins with
the incorporation of
formylmethionine, which is supplied
by a special “initiator” tRNA, called
tRNAMeti, which is recruited to the 30S
subunit by initiator factor IF-2.

The assembled 30S initiation complex


recruits the 70S (large) subunit to form
the functioning ribosome…

formylmethionine
7
221.S16 B. Keith
Prokaryotic translation: linear polysomes

8
221.S16
Translational initiation in eukaryotes: mRNA binding and scanning
(RBS)

scan->

9
221.S16
Translational initiation in eukaryotes: mRNA binding and scanning
The polypeptide chain is
begun with a methionine
attached to a special
initiator tRNAi, which is the
only tRNA that can bind the
ribosomes in the absence of
the 60S subunit…

A complex containing
initiation factors (eIF2, etc.),
the 40S subunit, and Met-
tRNAi binds the m7G 5’ cap, scan->
then scans along the mRNA
looking for the first AUG
sequence it can find…

(unlike bacteria, the initial


methionine in eukaryotes is
not formylated)

10
221.S16 B. Keith
Translational initiation in eukaryotes: mRNA binding and scanning

Once the 40S, Met-tRNAi


complex finds the first AUG
sequence, the 60S subunit
binds, leading to the release of
specific initiation factors (eg,
eIF2), and the start of protein
translation…

11
221.S16 B. Keith
Translational initiation in eukaryotes:
Two features recognized by ribosomes

Kozak sequence

12
221.S16
Translational termination: protein factors

When the ribosome encounters a stop


codon, specific release factors (RF1 and
RF2) enter the “A” site of the ribosome,
and bind directly to the mRNA. The
results in the hydrolysis of the bond
between the final amino acid and the
tRNA in the “P” site, releasing the newly
synthesized protein. RF3 promotes
dissociation of RF1 (or RF2), after which
the ribosome disassembles.

In bacteria, RF1 recognizes UAA and


UAG, RF 2 recognizes UAA and UGA. In
eukaryotes, there is only one Ribosomal
Release Factor (RRF), which recognizes
all three stop codons.

13
221.S16 B. Keith
Macromolecular mimicry: tRNA and Relaese Factors

tRNA
yeast RRF

anticodon
loop

14
221.S16 Selmer et al. Science (1999) 286:2349-2352
Properties of the Genetic Code

Non-overlapping code
No punctuation
3-nucleotide codons (triplet code)
Degenerate code
Start & stop codons
Nearly universal

15
221.S16
Genetic code

16
Mutations within coding regions

17
221.S16
Anatomy of an mRNA transcript

5’-untranslated region (leader) 3’-untranslated region (trailer)

H2N
COOH

[ORF: open reading frame]

18
[UTR: untranslated region]
221.S16
1 sequence and 3 reading frames

5’- CAT CAT CAT CAT CAT CAT CAT CAT


DNA
3’- GTC GTC GTC GTC GTC GTC GTC GTC

RNA 5’- CAU CAU CAU CAU CAU CAU CAU CAU frame 1

his his his his his his his his

5’- C AUC AUC AUC AUC AUC AUC AUC AU frame 2

ile ile ile ile ile ile ile

5’- CA UCA UCA UCA UCA UCA UCA UCA U frame 3

ser ser ser ser ser ser ser

19
221.S16
Translation: reading frames

20
221.S16 Figure 8-52a Molecular Biology of the Cell (© Garland Science 2008)
Translation: Open Reading Frames (ORFs)

vertical blue lines: stop codons


gray boxes: ORFs
red boxes: ORFs that begin with AUG codons
21
221.S16 Figure 8-52b Molecular Biology of the Cell (© Garland Science 2008)

Das könnte Ihnen auch gefallen