9.

Virus

Structured Questions
Question 1
In the 1950s, scientists were still debating on whether DNA or protein was the hereditary material.

Two biologists, Alfred Hershey and Martha Chase (1952), conducted two experiments using T2
bacteriophages and E. coli bacteria and proved conclusively that DNA was the hereditary material.

In the first experiment, Hershey and Chase labelled the DNA of T2 bacteriophages with radioactive
phosphorus-32 (32P). The 32P T2 bacteriophages were allowed to infect the E. coli bacteria. After a
fixed time, a blender was used to dislodge the T2 bacteriophages from the E. coli bacteria, and the
mixture was centrifuged to separate the bacterial cells from the phage particles left in the supernatant.
The amount of extracellular 32P in the supernatant was then measured.

In the second experiment, they labelled the protein of T2 bacteriophages with radioactive sulfur-35
(35S). The 35S T2 bacteriophages were allowed to infect the E. coli bacteria. As in experiment one, the
T2 bacteriophages were dislodged and the amount of extracellular 35S in the supernatant measured.

Fig. 1.1 shows the experimental setup.

The Hershey and Chase experiment

Fig. 1.1

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(a) (i) Explain why 32P was used to label DNA whereas 35S was used to label the protein. [2]
• Phosphorous is present as a component of the DNA nucleotide and not in any amino
acid;
• whereas S is found in the cysteine amino acid and not in DNA;

(ii) Explain how the experiment confirmed that DNA is the hereditary material. [3]
• [Idea of what hereditary material] Hereditary material carries genetic information that is
passed on from one generation to the next Or
In context – it refers to the material that carries genetic information for the production of
more viruses in the bacteria;
• Since 35S is found in the supernatant and not in the E. coli, indicating that capsid coat
did not enter into the cell/ remains outside the E. coli /bacteria;
• 32
P is not found in the supernatant/ only found in E. coli, indicating that DNA has been
injected into the cells and carries the genetic information for the production of the new
viruses;

(iii) Explain how the structure of the DNA allow for its replication in the E. coli bacteria. [3]
• DNA is double stranded and they are complementary to each other;
• Each can act as template for the synthesis of its complementary strand - semi-conservative
replication;
• Both strands are held by hydrogen bonds which can be hydrolysed by bacterial helicase;
• Helicase unwind the double-stranded DNA by breaking the hydrogen bonds between the
complementary bases;

(b) Describe the significance of named viral proteins to the reproduction of the T2 bacteriophages.
[3]
• Capsomeres form the capsid coat that protects viral DNA;
• Tail fibres and base plates bind to specific receptors on host cell;
• Contractile sheath contracts to drive a hollow tube in the tail into host cell, facilitating entry
of viral DNA into host cell;
• Enzymes eg. lysozymes are important to help to hydrolyse the bacterial cell wall;

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Question 2
In an experiment carried out, different concentrations of bacteriophage suspension were added to
the middle of four nutrient plates covered with bacterial lawn. The plates were incubated overnight
at 37oC. The observations were made and recorded as shown below:

Fig. 2.1

(a) Explain what caused the clear ring in plate 1. [3]

• The bacteriophage bind to specific receptors on the surface of the bacterial host cells and
inject their DNA into the cells;
• Enzymes coded by the viral DNA shut down the bacteria’s protein, RNA and DNA synthesis;
• And use the host protein synthesising machinery to replicate the viral DNA as well as
synthesise viral proteins to form new viruses;
• To release the new viruses, a phage-coded lysozyme breaks down the bacterial
peptidoglycan cell wall, resulting in cell lysis;
• Hence killing the bacteria, creating a bacteria free zone / clear ring;

(b) Account for the differences in observations from plate 1 to 3. [2]

• The size of the clear ring increases with the concentrations of bacteriophage added or
QV: Plate 1 has the smallest clear ring followed by plate 2 with plate 3 having the largest
clear ring;

• When more bacteriophages were added, more bacteria were infected and hence more
mature viruses produced;

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 • This allows for a greater number of bacteria being killed/lysed and hence a greater area of
clearance;

(c) What is the function of plate 4? [1]

• As a control to show that in the absence of bacteriophages, there is no clear ring formed ,
therefor the clear ring seen in plates 1-3 is due to the action of bacteriophage;

Question 3
Neuraminidase cleaves the sialic acid residues, on the surface of the infected host cell membranes,
to release new influenza viral particles. Oseltamivir (Tamiflu) is an antiviral drug that is effective
against the action of neuraminidase. The structures of sialic acid and oseltamivir are shown below.

Fig. 4.1

(a) (i) Using evidence from Fig 4.1, suggest the possible mode of action of oseltamivir. [3]
• Competitive inhibitor, Competes with sialic acid to bind to active site of neuraminidase;
• Structural resemblance between oseltamivir and sialic acid;
• Preventing / reducing cleavage of sialic acid thus preventing release of influenza viruses;

(ii) Suggest why antibiotics will be ineffective against viral infection. [2]
• Antibiotics kill living bacteria by targeting certain metabolic pathways eg. disrupt bacterial
peptidoglycan cell wall synthesis / inhibit protein synthesis in bacteria; [must have]

• [Idea of] Viruses are acellular /Viruses are obligate intracellular parasites and make use of
the host’s cellular machinery / A: named examples ribosomes, enzymes etc to replicate;
• Antibiotics used will kill the host cells instead/ Antibiotics have to kill host bacteria to prevent
viral infection;

The table shows an excerpt of the confirmed human cases of avian influenza reported by the World
Health Organisation (WHO).

Table: Cumulative number of confirmed human cases of avian influenza A (H5N1) reported to WHO
(19 June 2008)

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(b) Avian flu is typically described as “influenza caused by viruses adapted to birds”. Explain the
occurrence of human cases of avian influenza. [4]
• Caused by antigenic shift;
• Process by which 2 different strains of influenza virus infect a common host cell;
• When new viruses assembled, random assembly of RNA segments from the avian and
human influenza virus results in new combinations of RNA to form a new subtype
or
Antigenic shift may result when a human influenza virus acquires a new genome segment
from an avian influenza virus;

• genetic recombination / reassortment results in new subtype has a mixture of the surface
glycoproteins of the 2 original strains and hence can infect humans;
• The new genome segment causes a major change in the haemagglutinin which the human
immune system is not able to recognise;

(c) Describe 3 ways in which a virus may cause disease in animal host by damaging the infected
host cells. [3] [Any 3]
• Takes over host cell machinery and inhibits host cell DNA / RNA / protein synthesis;
• Causes cell lysis during release of new viruses;
• Causes death of many cells due to formation of syncytium;
• Insertion of viral glycoproteins at host cell membrane causes immune cells to kill infected
host cell;
• Causes release of inflammatory cytokines by infected host cell which in turn causes immune
cells to attack the host cell and neighbouring cells;

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Question 4
Fig. 4.1 shows how the human immunodeficiency virus (HIV) enters a macrophage, a type of white
blood cell.

Fig. 3.1

(a) Label the structures A, B, C and D.

A: Envelope / Phospholipid membrane; C: RNA;
Capsid coat / Capsomere /
B: D: Glycoprotein 120 / gp120;
Nucleocapsid;
[4]

(b) Explain why HIV is considered to be a retrovirus. [3]

• HIV carries a (+) single-stranded RNA as its genetic material/ genome;

• Which the virus use as a template to form a double stranded DNA via the enzyme reverse
transcriptase;
• which is inserted into the host DNA;
• Thus genetic information flows in the reverse direction, from RNA to DNA;

(c) Compare the entry process between the HIV and influenza virus. [3]
• Adsorption/ attachment of both viruses are by binding to specific cell surface receptors;

• Glycoprotein gp120 on the surface of the HIV binds to CD4, a cell-surface receptor found
on white blood cells/ T helper cells / macrophages of the host immune system;
• Haemagglutinin on the influenza viral membrane binds to sialic acid-containing receptors
on the host cell membrane;

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 • The influenza virus enters the host cell by receptor-mediated endocytosis;
• The HIV envelope does not enter via receptor-mediated endocytosis. Instead, the HIV
envelope fuses with the host cell membrane;

• Upon entry, the influenza virus forms an endosome / endocytic vesicle ;

• whereas the HIV virus does not form an endosome/ endocytic vesicle, the HIV releases
the viral contents into the host cell cytoplasm.;

(d) Upon completion of the entry process, describe how the genome of HIV is inherited. [3]

• RNA is reverse transcribed to complementary DNA strand by the enzyme reverse

transcriptase;
• The enzyme integrase catalyses the integration of the viral DNA into the host chromosome;
• The provirus genome is also replicated along with the host cell genome and all daughter
cells inherit the HIV genome;

(e) Suggest 2 ways in which retroviruses such as HIV can increase chances of a cancerous growth.
1. Insertion of viral DNA near promoter of proto-oncogene;
R! ‘active promoter’, as HIV does not carry proto-oncogene

2. Resulting in overexpression of proteins that stimulate cell division;

OR
3. Insertion of viral DNA within a tumour-suppressor gene;
4. resulting in loss-of-function mutation in genes coding for proteins which inhibit cell
division/ DNA repair proteins/ regulator of apoptosis/ proteins that control cell division are
non-functional;

[2]

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Essay Questions
Question 1
(a) Compare the ways in which lambda phage and human immunodeficiency virus reproduce
themselves. [10]
Features lambda phage
Viral structure used in • Attachment via tail fibres;
attachment
Type of receptor • Specific receptors;
Host cell • Bacterium;
Entry • Contraction of contractile
sheath drives a hollow tube
into host cell;
• Injection of viral DNA into
host cell;
Uncoating • Empty capsid coat left
outside host cell;
Viral genome • 1 copy of double-stranded
DNA;
What happen to the • May enter lysogenic cycle
viral genome where DNA inserts into host
DNA forming a prophage;
Synthesis of viral • Viral DNA exits host DNA
genome and proteins when induced;
• Viral DNA is replicated and
expressed to form viral
proteins;
Release • Phage-coded lysozyme
breaks down bacterial
peptidoglycan cell wall;
• New viruses released due to
host cell lysis;
• Both viruses reproduce in 5 main stages: attachment, entry, replication, assembly, release;
• Assembly of both viruses occurs in the cytoplasm;
• Both involve integration of viral DNA into host genome;
Question 2
(b) Discuss the importance of membranes in the reproductive cycle of influenza virus and HIV. [7]
human immunodeficiency virus
• Attachment via gp120;
• CD4 receptors;
• Human T-helper cell;
• Fusion of viral envelop with host
cell membrane;
• Release nucleocapsid into
cytoplasm;
• Capsid coat is enzymatically
hydrolysed to release viral RNA
into cytoplasm;
• 2 copies of single-stranded
RNA;
• Reverse transcriptase
synthesises double-stranded
DNA from viral RNA;
• Viral DNA inserts into host DNA,
catalysed by integrase, forming
a provirus;
• Viral DNA is transcribed when
activated;
• mRNA serves as viral genome
and template to form viral
proteins;
• Viral glycoproteins inserted into
host cell membrane;
• New viruses bud off host cell at
the exit point;
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 • Host cell membrane contains specific receptors;
• Viral envelop contains specific viral glycoproteins which bind to these specific receptors for
viral attachment to host cells;

• Influenza virus binds to specific receptors containing sialic acid on respiratory epithelial
cells;
 Via haemagglutinin;
• Invagination of host cell membrane allows entry of influenza virus via endocytosis;
 Influenza virus found in endosome;
 Fusion of viral envelop with endosome membrane releases nucleocapsid into the
cytoplasm;
 Uncoating releases viral RNA, accessory proteins and RNA-dependent RNA
polymerase into the cytoplasm to allow for replication;

• HIV binds to CD4 receptors on T-helper cells;

 Via gp120;
• Fusion of viral envelop with host cell membrane releases nucleocapsid into the
cytoplasm;
 Uncoating releases viral RNA, reverse transcriptase and integrase into the cytoplasm
to allow for replication;

• Endomembrane system including rough endoplasmic reticulum and Golgi apparatus

are involved in the synthesis, modification and transport of viral glycoproteins;
• Viral glycoproteins are inserted into host cell membrane at exit points;
• Assembled nucleocapsids move to the exit points and new viruses are released via
budding;
 Involving the evagination of host cell membrane to surround the nucleocapsid;
 New viruses thus acquire the viral envelop with viral glycoproteins;

Question 2 (2016 VJC Promo Q7)

Coherent sequential use of cellular structures to support virus reproduction

7 Explain how the human lung epithelial cell is able to support the reproduction of [13]
Influenza virus.
Answer from the perspective of the host cell

Max 2m for points in introduction that sets the framework for structure-function link of host cell organelles being
exploited by virus
**Links must be made between the cell structure and the stage of virus reproductive cycle to gain full marks
*repeating points to be awarded once;
*students are required to address this question from the perspective of how cell structures are subsumed into
the mode of replication of Influenza virus, regurgitation of the five stages will not be awarded full marks

The BONUS mark is awarded for quality of answer, when:

i. Student’s essay includes at least FOUR different parts of the cell - cell surface membrane, nucleus,
RER, Golgi body, etc. [including ribosomes, secretory vesicles, transport vesicles, endosome; Also
accept cytoplasm, nuclear pore] to reflect structure-function relationships of cell organelles of the host
cell are exploited by the virus to support its mode of reproduction.

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 ii. Student is able to connect virus reproductive stages with role of host organelles in a coherent manner,
even if essay is not completed.

Setting the context (2 max)

1. Influenza virus reproduces in five basic stages – attachment, entry, replication, assembly and
release (identify the stages of virus replication); [Award only once for the stages]
2. The human lung epithelial cell is the specific target cell of Influenza virus which makes use of
host cell machinery to reproduce to multiply its numbers to infect successive rounds of host
cells/ idea of why virus is an obligate intracellular parasite;

Cell Surface Membrane (CSM) (3 max)

– has specific integral glycoproteins that contains sialic acids which interact with viral
haemagglutinin (HA) glycoprotein during ATTACHMENT stage;
• Fluidity of the membrane allows for invagination to form endosome around virus to
facilitate its entry into cell by endocytosis during ENTRY stage;
• Low endosome pH facilitates fusion with viral envelope as well as trigger uncoating of
viral genome from viral structural components for entry into host cytoplasm;
• Contains incorporated viral HA and NA at exit points where RELEASE by budding of
mature virus progeny occurs new viruses acquire viral envelope in the process of
budding ► next cycle of infection of host cells

Nuclear envelope/ nucleus (2 mx)

• Site where genome replication of viral RNA occurs using free ribonucleotides and viral
RNA polymerase;
• contains nuclear pores that allow for transport/ passage of RNA-dependent RNA
polymerase complex and viral genome from cytoplasm during REPLICATION stage;
• nuclear pores – allow for the transport of capsomeres from cytoplasm into nucleus for
assembly/maturation stage [award once only]a

Cytoplasm (6 max)
• contains free ribosomes where synthesis of viral proteins eg. capsid proteins/
structural proteins and enzymes;
• ► synthesised capsomeres move back to nucleus to wrap around RNA genome to
form nucleocapsid during ASSEMBLY/ MATURATION stage ► back to cytoplasm (to
assemble with rest of structural components);

• contains rough ER (attached ribosomes) where synthesis of glycoproteins HA and NA

targeted for insertion into the CSM;
• ► RER membrane bud off into transport vesicles carrying HA and NA to move to cis face
of GA for membrane fusion;;

• contains Golgi apparatus which receives transport vesicles from RER (at their cis face)
► site of where glycosylation of HA and NA occurs, sorting and packaging into
secretory vesicles (at trans face);
• ► secretory vesicles with membrane containing inserted HA and NA move to CSM ►
fuse with CSM at specific sites called exit points;
• ► HA and NA are now incorporated into host plasma membrane at exit points for
RELEASE/ EXIT/ BUDDING;

Examiner’s Comments:
• There were several elegant essays that scored full marks without the additional well-deserved Bonus
mark for quality writing. These students demonstrate the skill of understanding the requirements of the

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 question, answering from the perspective of the host cell and provide a coherent picture of how each
cell structure supports the sequence of virus reproduction.
• Several students provided detailed drawings of the stages of virus reproduction in a host cell. As these
students have also provided the same details in their text, no extra marks were awarded to their drawing.
Marks will be awarded to supplementary details in the drawings that were not addressed in the text.
• Several students fail to contextualise their responses from the perspective of the host cellular structures
and launch into a regurgitated reproductive cycle of the influenza virus, showing that they have yet to
master the skill of unpacking the question. These students will not be awarded full marks (as they will
not attain the Bonus mark even if they satisfy the criterion of listing at least four cellular structures in
their essay.
• The Bonus mark is awarded to incomplete essays that answer from the perspective of the host cell and
have identified at least four cellular structures in a coherently sequential process of virus reproduction.
• Fundamental misconceptions surfaced in some scripts showing that students memorised without
understanding and confused Influenza virus with HIV, in terms of surface glycoproteins (e.g. gp120
instead of HA), enzymes (e.g. reverse transcriptase instead of RNA-dependent RNA polymerase).
• Some students demonstrate a poor understanding of biological terms, confusing the term ‘exocytosis’
with ‘budding’ during the release phase of virus,
• Many students fail to use the precise term ‘cell surface membrane’ when referring to specific receptors
on the host cell surface that are recognised and bound by HA. It is important to understand that a cell
has numerous receptors located not just on its CSM, but elsewhere inside the cell, e.g. cytoplasmic
receptors. These are intracellular receptors and include steroid receptors.
• Students use the term ‘cell membrane’ loosely. Cell membrane can mean membrane enclosing
organelles. Cell surface membrane (CSM) refers to the membrane enclosing the cell. It can also be
referred to as the plasma membrane.
• Similarly, the receptor that is recognised by HA should be accurately called specific as many types of
receptors contain sialic acid (a type of sugar) are found on the surface of a single cell. By saying specific,
it clarifies that of all the different types of sialic-acid containing receptors found on the CSM of a cell,
there is one specific type that is recognised by the HA of Influenza virus.

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