65.
Viruses that cause cancer
Summary
A causal link with viruses is found in about 15% of all human tumors. Tumor-initiating viruses
cause malignant cell transformation in vitro and can induce tumor formation in vivo (the latter
in interaction with additional genetic and environmental factors). Tumor viruses are either
directly acting (if they have a tumor-causing oncogene in their genome) or indirectly acting
(these increase the chance of tumor formation in the absence of such a gene by causing chronic
inflammation). The genetic material of viruses is either DNA or RNA. During tumor formation,
the general rule is that the viral genome must be integrated into the genome of the infected host
cell in order to produce a stable cell transformation or a tumor cell. Although many people are
infected with tumor viruses, only a few actually develop tumors (the mechanism of this process
is similar to that of nonviral tumor development). Tumor viruses and their infection of cells are
also a good experimental model system for studying tumor formation (e.g. SV40 infecting
monkey cells in the wild, or polyoma virus infecting mouse cells).
Keywords: DNA tumor virus, early and late DNA regions and proteins, hepatitis B virus,
human papilloma virus, cervix carcinoma, Epstein-Barr virus, Burkitt lymphoma, human
herpesvirus 8, Kaposi’s sarcoma, RNA tumor virus, strongly and weakly transforming
retroviruses, Rous sarcoma virus (RSV), reverse transcription, long terminal repeat (LTR)
sequence, retroviral oncogene, v-src, cellular protooncogene, c-src
DNA tumor viruses
The obligatory components of a DNA virus particle (virion) are the genetic material (in their
case DNA) and a surrounding cover of viral proteins called capsid. The entire DNA of a virus
can be divided into two main regions. The early regions, expressed during the early stages of
virus infection (in about the first 24 hours), and the late regions (or genes), transcribed after this
time. The early region does not encode structural elements of the virus particle, but so-called
early proteins. These are used to prepare the infected cell for replication of the virus. The early
DNA region is transcribed into a primary transcript in the infected host cell, from which mature
mRNAs are generated by alternative splicing. These are translated to form the early proteins
required for viral replication, the best known of which are:
- the large T-protein, which stimulates DNA replication in the host cell nucleus, and
regulates transcription and immortalizes the cell,
- the medium T protein (encoded only by the genome of certain viruses, e.g. polyoma
virus): it is located in the plasma membrane of the infected cell and causes
transformation of the cell's morphology,
- small t-protein: it is found in the cytoplasm and completes the transformation of the
infected cell.
The genes in the late region of the viral DNA encode the structural protein elements of the virus
particle (in other words, those of the virion).
In human tumors, the viral DNA is present in the host cell in (latent) extrachromosomal form
for a period of time. This allows the virus to replicate successfully. If, however, the viral DNA
becomes integrated into the genome of the host cell, the increased expression of early viral
genes (which regulate replication) in the new DNA environment transforms the host cell and its
progeny into tumor cells. It is from these cells that the tumor develops.
DNA viruses that cause human tumors
• Hepatitis B virus (HBV)
Infects liver cells → acute inflammation of the liver / hepatitis (with a healthy immune system
it can be defeated over time, while with a weakened immune system the virus can persistently
colonize liver cells) → this condition is a transition into chronic hepatitis. In the latter case,
liver tissue is destroyed but tries to regenerate → the liver tissue structure is remodeled / liver
cirrhosis → the chances of developing liver cancer are multiplied.
The development of liver cancer is associated with chronic inflammation, the exact molecular
genetic mechanism is not known.
• Human papillomavirus (HPV)
This is a family of viruses with > 100 different virus types. It infects epithelial cells and can
cause a wide range of tumors from benign warts (where typically less early protein is expressed
in the cells of the superficial layers of the epithelium) to malignant tumors affecting the
anogenital region. The best-known tumor type of the latter is human cervical carcinoma:
This is a sexually transmitted disease. The virus enters the cells of the deeper layers of the
epithelium through small lesions and then binds to cell surface integrins and is internalized by
endocytosis. The genetic material of the virus is incorporated into the genome of the host cell
which can be detected (> 99%) in the deeper / basal layers of the infected epithelium, in the
tumor cells there. A long, multiyear transition from infection to malignant cell proliferation is
possible. Some conditions may sometimes become spontaneously normalized.
During tumor development, the so-called E6 and E7 early viral proteins are overproduced and
interfere with proteins (p53 and Rb) that regulate the cell cycle and apoptosis. Viral
oncoproteins also interfere with normal duplication of centrosomes → common chromosomal
abnormalities in tumor cells.
• Herpesviruses
This is another large family of viruses, of which the two best known tumor-causing members
are Epstein-Barr virus and human herpesvirus 8.
• Epstein-Barr virus (EBV)
It has been named after its discoverers. It infects many (~90% of us will get it at some point in
our lives, even if asymptomatically). In individuals with a healthy immune system, usually →
infectious mononucleosis, while in people with a weakened immune system (caused e.g. by
malaria or along with AIDS) → Burkitt lymphoma, or nasopharyngeal tumor.
An important factor in the molecular mechanism of tumor development may be virus-induced
B-cell hyperproliferation → resulting in characteristic abnormalities (so-called reciprocal
translocation) in the structure of certain chromosomes.
• Human herpesvirus 8 (HHV8, formerly known as Kaposi's sarcoma-associated
herpesvirus = KSAHV)
HHV-8 in healthy individuals → an asymptomatic infection without underlying disease →
in the final stage of AIDS Kaposi's sarcoma develops upon infection = skin lesions / vascular
tumors of the subcutaneous connective tissue (indicator disease!)
Molecular mechanism of tumor development:
- the viral genome is detectable in all tumor cells
- the viral genome contains transforming genes the protein products of which are:
o angiogenic factor → strong tumor vascularization
o cyclin-like factor → rapid cell division
o anti-apoptotic / Bcl2-like factor → tumor cell immortalization
RNA tumor viruses
Their most important representatives belong to the group of retroviruses. They rewrite their
genetic material, consisting of RNA, into DNA by means of their reverse transcriptase enzyme
when the host cell is infected.
The major components of retroviral particles are (Fig. 1.):
• they are covered by a phospholipid bilayer (envelope), in which the Env proteins
(encoded by the env sequence of the genome) are embedded
• the nucleocapsid contents within consists of: genomic RNA molecules, Gag proteins
(encoded by the gag sequence of the genome), reverse transcriptase enzyme (encoded
by the pol sequence of the genome)
phospholipid
bilayer
Env protein
Gag protein
reverse
transcriptase
viral RNAs
Figure 1. The structure of retroviruses.
The main steps in viral infection (Fig. 2.): viral attachment to the host cell’s surface → viral
phospholipid bilayer fuses with the host cell’s plasma membrane → viral capsid enters the
cytoplasm and becomes unpacked → the released viral genome RNA is reverse transcribed
back first into single- and then into double-stranded DNA (at the ends of which are the long
terminal repeat = LTR sequences) → the double-stranded DNA is translocated to the host cell
nucleus and then randomly inserted somewhere in the host cell genome (thus becoming a so-
called "provirus"). Virus-specific RNAs are formed from the integrated viral genome (some of
which serve as the genome of progeny viruses, others as templates for translation of viral
proteins). The virus-specific RNAs are transported into the cytoplasm and the viral protein
encoders are translated → the virus-specific genomic RNAs and newly formed viral proteins
assemble into new viral particles and are then released from the host cell by budding / sprouting
cell membrane
membrane fusion
virion
tRNA primer
envelope
protein nucleo- virus RNA
capsid
reverse transcription
double-stranded cDNA
LTR LTR
nuclear envelope
capsid integration
proteins
cellular genome
provirus
transcription, splicing
virus genomic RNA genomic RNA
budding
mRNA
translation
vesicular transport
of Env protein endoplasmic
reticulum
Figure 2. The infectious cycle of retroviruses.
The infection usually does not kill the host cell (it is nonlytic) and the infected cell that produces
viruses can become transformed. If the infection affects the germ cell line during early
development → the formation of so-called endogenous retroviruses is possible (these are not
infectious and their genes are usually not transcribed, but if their expression is activated → they
may contribute to the development of autoimmune diseases /e.g. SLE or Sjögren's syndrome/.)
Major human retroviruses of medical significance
• HTLV-1 = Human T-cell lymphotropic virus → adult T-cell leukemia / lymphoma (more
common / Japan, Caribbean, Africa)
The tax regulatory protein coded by the viral genome may be important in the molecular
mechanism of tumor development / it influences several genes in the host cell that are
also regulated by growth factors.
• HIV = Human Immunodeficiency Virus (formerly HTLV-3) is not primarily a tumor
virus but a causative agent of AIDS (AIDS = Acquired Immune Deficiency Syndrome:
in the course of the disease, however, the development of secondary tumors is common,
e.g. Kaposi's sarcoma, or lymphomas due to weakened immunity caused by destroyed
helper T-cells!)
Historical significance of retroviral tumors in various species (vertebrates)
Cell-transforming effect of retroviruses
• Strongly / rapidly transforming retroviruses: e.g. Rous sarcoma virus (RSV)
o rapid tumor initiation (1-2 weeks) in vivo
o polyclonal tumor (e.g. sarcoma in chicken)
o in vitro (cell culture) virus replication and cell transformation
o oncogene in the viral genome (v-src)
o viral genome size ~ 10 kb
• Weakly transforming retroviruses: e.g. avian leukosis / leukemia virus (ALV)
o slow tumorigenesis (only months later) in vivo → monoclonal tumor (e.g.
lymphoma in birds)
o there is also an RSV variant with similar behavior and structure, the so-called
transformation defective / deletional RSV (td-RSV)
o in vitro (cell culture) viral replication but no cell transformation
o no oncogene in the viral genome
o viral genome size ~8.5 kb (1.5 kilobases shorter than that of strongly oncogenic
/ transforming retroviruses and this difference in length is equivalent to about
the size of a protein coding gene)
Identification of the first retroviral oncogene
Normal / wild-type RSV genomic RNA (~10 kilobases in length) → reverse transcribed back
to radioactive cDNA → cDNA fragmentation into smaller pieces and hybridization with td-
RSV genomic RNA (~8. 5 kilobases) from the resulting mixture a single-stranded cDNA ~ 1.5
kilobases long can be isolated by hydroxyapatite chromatography → v-src (the first successfully
identified retroviral oncogenic sequence, the viral equivalent of the c-src cellular
protooncogenic sequence) (Fig. 3.)
Figure 3. The identification of the v-src retroviral oncogene.
Other retroviral oncogenic sequences (only the name of the oncogene and its function is
important)
Virus-induced Function of
Oncogene Host
tumor oncoprotein
Non-receptor tyrosine
src Chicken Sarcoma
protein kinase
myc Chicken Myelocytoma etc. Transcription factor
erbA Chicken Erythroleukemia etc. Transcription factor
Receptor tyrosine
erbB Chicken Erythroleukemia etc.
protein kinase
jun Chicken Fibrosarcoma Transcription factor
fos Mouse Osteosarcoma Transcription factor
Non-receptor tyrosine
abl Mouse Leukemia, sarcoma
protein kinase
raf Mouse Sarcoma MAPKKK
ras Rat Sarcoma, etc. Monomer G-protein
sis Monkey Sarcoma PDGF
akt Mouse Lymphoma Protein kinase
Receptor tyrosine
kit Cat Sarcoma
protein kinase
Significance of the discovery of viral oncogenes
Their cellular homologues, the cellular protooncogenes, are highly conserved / found in most
eukaryotic species. Their encoded products are important regulators of cell division, growth,
survival, and differentiation.
During their evolution, tumorigenic retroviruses have "captured" their viral oncogenes by
infecting their host cells (oncogene capture). All viral oncogenes have cellular counterparts, but
the reverse is not true.
The main steps in the process of oncogene capture are: integration of a provirus without
oncogene into the host cell genome, near a healthy / normal protooncogene → deletion →
transcription and splicing → the genomic retroviral RNA formed recombines with helper virus
RNA (Fig. 4.).
provirus of Moloney exons of c-abl integration
virus
cellular
DNA
LTR gag pol env LTR
deletion
gag
transcription
splicing
5` 3`
gag v-abl
5` 3`
retrovirus RNA
recombination
5` 3` transducing
gag v-abl retrovirus
Figure 4. Steps of the formation of a transducing retrovirus.
Why do viral oncogenes cause tumors while their cellular protooncogene counterparts do not?
Cellular protooncogenes: function in a normal regulatory environment, contain introns and are
not mutated → their expressed protein product’s function is normal / well regulated.
Retroviral oncogenes are transcribed under the control of LTR (a very strong promoter!), lack
introns and are often loaded by point mutation(s), truncation(s), or gene fusion(s) → their
protein products are constitutively / hyperactively functioning mutants.
Questions
Name two DNA viruses that can cause human tumors!
hepatitis B virus, Epstein-Barr virus, human papilloma virus, etc.
What happens to the genome of oncogenic DNA viruses during malignant transformation of
cells?
integrates into the genome of the host cell
Name two oncogenic DNA viruses!
SV40, papilloma virus, adenovirus, etc.
Name the cell in which a DNA virus causes lytic infection!
permissive cell
Which virus has a role in the development of human cervical carcinoma?
human papillomavirus
Which virus has a role in the development of Burkitt lymphoma?
Epstein-Barr virus
Name the DNA copy of the retroviral genome integrated into the host cell’s genome!
provirus
Name the enzyme that synthesizes the retroviral provirus!
reverse transcriptase
Name the genes of a weakly oncogenic retrovirus (3)!
gag, pol, env
What is v-src? Where is it found?
the retroviral oncogene of Rous sarcoma virus
What is c-src?
a proto-oncogene/cellular oncogene homologous to v-src
What does the abbreviation LTR stand for?
long terminal repeat
Name two retroviral oncogenes!
v-src, v-myc, v-jun, etc.
Give an example for a retroviral oncogene lacking a proto-oncogenic equivalent!
no such is known
