1.

Explain the effect of the DELETION OF the following sequences or features on mRNA.

Please give the answer according to the correct question.

(a)

the shine Dalgarno sequence (4 marks)

The Shine-Dalgarno (SD) sequence is a ribosomal binding site in

prokaryotic messenger RNA, generally located around 8
bases upstream of the start codon AUG. Prokaryotic mRNAs with
Shine-Dalgarno sequence deleted are either not translated or are
translated inefficiently.
(b)

AAUAAA consensus sequence (4 marks)

The deletion of the AAUAA consensus sequence will prevent binding

of the cleavage and polydenylation factor (CPSF), thus resulting in
no cleavage or polyadenylation of the pre-mRNA. This would affect
the stability and translation of the mRNA.
(c)

5 cap (4 marks)

The deletion of the 5 cap would most likely prevent splicing of the
intron that is nearest to the 5 cap. Ultimately, elimination of the
cap will affect the stability of the pre-mRNA as well as its ability to
be translated.

2.
Fill in the blanks and give the functions for each protein factors involved during protein
synthesis in prokaryotes and eukaryotes. (9 marks)

Protein factors

Function (s) (2 marks each)

(1 mark each)
Prokaryotes
IF1

Eukaryotes

eIF1A

IF2

eIF4A

IF1 Prevents tRNA from binding A sites on

30S
eIF1A binds 40S at A site
IF2 facilitates binding of fMet-tRNAfMet
eIF4A act as ATPase-dependent RNA
helicase

EF-Tu

eIF1a

EF-Tu helps aminoacyl-tRNA move onto a

free site on the ribosome. In
the cytoplasm, EF-Tu binds an
aminoacylated, or charged, tRNA
molecule.
eIF1a carries incoming aminoacyl-tRNA to
the A site

3.
Secreted proteins typically interact with two membrane-bound organelles prior to secretion.
Describe the activities of the two organelles in terms of preparing the polypeptide for secretion out of
the cell.

Pathway taken by secreted proteins ( the secretory pathway) :

Rough Endoplasmic Reticulum (RER) Golgi apparatus Secretory
vesicles Cell exterior.
Once the ribosomes synthesizing, these proteins become bound to the
RER, the proteins enter or cross the RER membrane . Secreted proteins,
including hormones, interact with rough endoplasmic reticulum (RER) and
Golgi apparatus (or lysosome or plasma membrane). If a newly formed
polypeptide contains a signal sequence, a short stretch of amino acids
with instructions for its destination in the organism, will bind to a signal
recognition particle. Then both bind to a receptor protein in the
membrane of the RER, and translation proceeds. The signal sequence is
removed by an enzyme in the lumen of RER. The polypeptide continues to
elongate until translation terminates. The ribosome is then released, and
the protein folds inside the RER. Next the protein may move elsewhere
within the endomembrane system, such as the Golgi apparatus (or
lysosome or plasma membrane). If it lacks specific signals for destination
within the endomembrane system, it will usually be secreted from the
vesicles that fuse with the plasma membrane.
(8 marks)

4. How much energy is required to translate a protein that is 20 amino acids long? Include in your
analysis the amount of energy to add the amino acids to the tRNAs, and all of the steps in
translation. Show your work for any partial credit.

80 energy is required to translate a protein with 20 amino acids long.

Amino acid activation : ATP AMP + PPi 2Pi
:
20 x 2 = 40
Initiation
: GTP GDP + Pi
:
1=1
Elongation
: GTP GDP + Pi ( elongation )
:
19 x 2 =
38
: GTP GDP + Pi ( translocation)

Termination
Total
energy

: GTP GDP + Pi
:

:
:

1=1
80

(6 marks)