Tumor Viruses

For most viruses:

Replication Lysis Progeny virions

Genome all viral proteins

Lytic Life Cycle

Non-structural and structural proteins made

1
 Tumor Viruses
Virus

Latent Life Cycle Cell

Integration (often)

Transformation

Some virus-specific proteins expressed (early functions) - No

mature virus
Viral structural proteins are not expressed
Changes in the properties of host cell - TRANSFORMATION
Sometimes latency may terminate – cell must be infected by
2
complete virus
 Tumor Viruses
Transformation:
• Loss of growth control
• Reduced adhesion
• Motility
• Invasion
• Ability to form tumors - viral genes interfere with control of
cell replication and other aspects of the cell phenotype

• Transformed cells frequently exhibit chromosomal

aberrations
3
 Tumor Viruses
TRANSFORMATION

VIRAL TRANSFORMATION
The changes in the biological functions of a cell that result from
REGULATION
of the cell’s metabolism by viral genes and that confer on the
infected cell certain properties characteristic of
NEOPLASIA

4
 Tumor Viruses
• Both DNA and RNA tumor viruses can
transform cells

• Integration of viral genome into the host

chromosomes often occurs

• Similar mechanisms of transformation by

each type of tumor virus
5
Two Major Classes of Tumor Viruses
DNA Tumor Viruses
DNA viral genome
DNA-dependent
Host RNA
DNA polymerase
polymerase
(Host or viral)
Viral mRNA

Similar to host cell!

Viral protein

6
 RNA Tumor Viruses
Viral RNA genome
Reverse transcriptase (Virus-encoded)

Viral DNA genome (integrated)

DNA-dependent RNA polymerase (Host
IMPORTANT RNA pol II)

Viral genomic RNA

Splicing (Host splicing enzymes)

messenger RNA
Important: Use HOST
RNA polymerase
viral protein to make its genome
An enzyme that
normally
Virus
makes mRNA 7
 DNA Tumor Viruses
DNA genome
Host RNA
polymerase II
mRNA
Host enzymes

protein

virus

OR TRANSFORMATION
In transformation usually only EARLY functions are expressed
8
 DNA Tumor Viruses In
Human Cancer

Papilloma Viruses
• cause natural cancers in animals
• cause benign warts
• ubiquitous
• epitheliotropic - most human tumors are malignancies of epithelial
cells
9
 DNA Tumor Viruses In
Human Cancer

Papilloma Viruses

• Epidermodysplasia verruciformis

wart malignant
skin squamous cell carcinoma

10
DNA Tumor Viruses In
Human Cancer
Epidermodysplasia
verruciformis

Papilloma virus

© 2008
Dermatology
Online Journal

11
 DNA Tumor Viruses In
Human Cancer
Papilloma Viruses
urogenital cancer
wart malignant squamous cell carcinoma

Squamous cell carcinoma:

Larynx
Esophagus All histologically similar
Lung

10% of human cancers may be HPV-linked 12

 DNA Tumor Viruses In
Human Cancer
Papilloma Viruses
• >100 types identified - most common are types 6 and 11
• Most cervical, vulvar and penile cancers are ASSOCIATED with
types 16 and 18 (70% of penile cancers)

EPIDEMIOLOGIAL STUDIES BUT:

HPV 16 and HPV 18 do transform human keratinocytes

Effective Vaccine
(quadrivalent recombinant HPV 6, 11, 16 and 18 proteins made in
yeast - Gardasil) 13
 Papilloma Viruses
• The important transforming genes in
papilloma viruses are: E6 and E7
• Early genes - Not encoding structural
proteins
• Oncogenes

14
 DNA Tumor Viruses In
Human Cancer
Polyoma Viruses
• Simian virus 40 - juvenile hamster sarcomas, transformation
• Polyoma - mouse leukemia, in vitro transformation
• Human polyomas (JC and BK) - monkey sarcoma, transformation
Possible association of BK with human prostate cancer

Polyoma virus transforms cells when the genome is incomplete

Early functions are necessary - ONCOGENES

15
JC: PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY (PML)
DNA Tumor Viruses In
Human Cancer
Adenoviruses
Highly oncogenic in animals
Only part of virus integrated

Always the same part

Early functions

E1A region: 2 T antigens

E1B region: 1 T antigen
E1A and E1B = Oncogenes 16
 DNA Tumor Viruses In
Human Cancer
Common pattern

Early functions (non-structural) proteins are involved

in transformation

• Papilloma: E6 and E7
• Polyoma: Large T and small T antigen
• Adenovirus: E1A and E1B

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 DNA Tumor Viruses In
Human Cancer
ONCOGENE
A gene that codes for a protein that potentially can transform
a normal cell into a malignant cell
An oncogene may be transmitted by a virus in which case it is
known as a VIRAL ONCOGENE

v-onc
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 DNA Tumor Viruses In
Human Cancer
Herpes Viruses
Considerable evidence for role in human cancer
• Some very tumorigenic in animals
• Integrated viral DNA found in small proportion of tumor cells:
“hit and run”

19
 DNA Tumor Viruses In
Human Cancer
Epstein-Barr Virus
• Burkitt’s Lymphoma
• Nasopharyngeal cancer
• Infectious mononucleosis (glandular fever)
• Transforms human B-lymphocytes in vitro

• Burkitt’s lymphoma: malarial infested regions

• Nasopharyngeal cancer: China, SE Asia – diet?
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DNA Tumor Viruses In
Human Cancer
Human herpes virus – 8
Kaposi’s Sarcoma Herpes Virus
Kaposi’s sarcoma

Hematologic malignancies

• Primary effusion lymphoma

• Multicentric Castleman's disease (MCD) – a rare
lymphoproliferative disorder (AIDS)
• MCD-related immunoblastic/plasmablastic
lymphoma
• Various atypical lymphoproliferative disorders
21
DNA Tumor Viruses In
Human Cancer
Hepatitis B Virus
DNA genome
RNA polymerase II Host enzyme

RNA Provirus
Reverse transcriptase
Viral enzyme
DNA genome

22
 DNA Tumor Viruses In
Human Cancer
Hepatitis B continued

• Vast public health problem

• 10% of population in underdeveloped countries are chronic carriers

•Long latency

23
 DNA Tumor Viruses In
Human Cancer
Hepatitis B continued

Epidemiology:
• Strong correlation between
HBV and hepatocellular
carcinoma
• China: 500,000 - 1 million new
cases of hepatocellular carcinoma
per year
• Taiwan: Relative risk of getting
HCC is 217 x risk of non-carriers
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 DNA Tumor Viruses In
Human Cancer
Summary

• These viruses can transform cells or have lytic life cycle

• Often integrate into host genome

• In transformation often ONLY early genes

(non-structural) are transcribed
• These are genes that are also necessary for a
PRODUCTIVE infection
• True viral genes
25
 RNA Tumor Viruses
RNA Genome - Retroviruses
RNA-dependent DNA Polymerase encoded by virus
REVERSE TRANSCRIPTASE
RNA genome
Reverse transcriptase virus

DNA genome
Integrase virus

Integrates
Host RNA polymerase II host

RNA genome 26
RNA Tumor
Viruses

27
RNA Tumor Viruses
A normal retrovirus has:
3 genes
GAG : internal proteins
ENV: Envelope glycoproteins
POL: Enzymes
Reverse transcriptase – RNase H
Integrase
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Protease
 RNA Tumor Viruses
RNA is:
• Diploid Capped and polyadenylated
• Positive sense (same as mRNA)
Viral RNA cannot be read as mRNA
(even though same sense)
New mRNA must be made
Virus must make negative sense DNA before proteins are
made
Therefore virus must carry REVERSE TRANSCRIPTASE
29
into the cell
RNA Tumor Viruses

30
 RNA Tumor Viruses
Groups of Retroviruses
• Oncovirinae important

Tumor viruses and similar

• Lentiviruses important

Long latent period

Progressive chronic disease
Visna HIV
31
 RNA Tumor Viruses
Retroviruses known to cause human cancer
• Human T cell lymphotropic virus -1 (HTLV-1)
Adult T cell leukemia, Sezary T-cell leukemia
Africa, Caribbean
S. America (Peru, Bolivia)
Some Japanese Islands
Okinawa, Kiyushu, Shikoku (12 - 16% infection rate)

32
 RNA Tumor Viruses
Human T cell lymphotropic virus -1 (HTLV-1)
UNITED STATES AND OTHER WESTERN COUNTRIES
IV DRUG USERS
US rate of infection about one tenth of that of HIV
BUT half as prevalent as HIV in IV drug users

Also causes: Tropical spastic paraparesis

• (affects the gray and white matter of the spinal
cord - myelopathy)
• 1-4% of infected people
Immunosuppression 33
 RNA Tumor Viruses
Retroviruses known to cause human cancer
• Human T cell lymphotropic virus -2 (HTLV-2)

Hairy cell leukemia

Americas, particularly in native American populations
New Mexico (Navajo and Pueblo Indians)
Florida (Seminole Indians)
Seroprevalence in these populations > 20%
Women over 50: seroprevalence - up to 50% in some populations

• HIV ? 34
 RNA Tumor Viruses
Retrovirus Life Cycle
Bind to Endocytosis
surface receptor

Fusion of membranes

Release of nucleocapsid to cytoplasm

Nucleus
35
 RNA Tumor Viruses
Parental RNA
Reverse transcriptase

RNA/DNA Hybrid
Reverse
transcriptase
Linear DNA/DNA duplex

Circular Duplex DNA

Integrase Host DNA
polymerase
Integration Replication (DNA genome in cell)
Host splicing
Host RNA pol II enzymes
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Transcription Viral RNA genome mRNA protein
 RNA Tumor Viruses
Drawback to this lifestyle

Genomic RNA
Reverse transcriptase

DNA
Host RNA pol II
Genomic RNA

Pol II is a host enzyme that, in the uninfected cell, makes mRNA

When making mRNA, pol II does not copy entire gene to RNA
37
 Problem of using RNA pol II to copy a gene

RT Viral
primer genomicRNA
Reverse
transcriptase
dsDNA

RNA synthesis
initiation site

promotor

RNA pol II
RNA synthesis termination
site

38
Result: New copy of viral RNA is shorter - lacks control sequences
 RNA Tumor Viruses
RNA polymerase II will not copy
Upstream sequences from transcription initiation site
• Promotors / Enhancers
Down stream sequences from transcription termination site
• Enhancers / Poly A site / termination site

?
Perhaps virus could integrate downstream of a promotor etc so
that the cell provides sequences

OR
Virus provides its own promotors etc
39
BUT not copied!
 RNA Tumor Viruses
Repeat Repeat
region region
Clue: Difference in the two forms
RNA

R U5 GAG POL ENV U3 R

DNA

U3 R U5 GAG POL ENV U3 R U5

LTR LTR 40
 R U5 Viral RNA U3 R

Reverse
transcriptase

U3 R U5 U3 R U5
Long terminal repeats are formed
promotor
POLII POLII

RNA initiation site RNA termination site

41
Retroviruses can have only one
promotor region
Contained in U3

LTR LTR

POLII POLII

RNA initiation site RNA termination site

Therefore only one long RNA can be made

U5
Therefore mRNA requires processing
Explains why RNA has to be positive sense 42
 Some retroviruses have an
extra gene

“typical retrovirus”

R U5 GAG POL ENV U3 R

Rous Sarcoma Virus

R U5 GAG POL ENV SRC U3 R

43
Some retroviruses have an oncogene
instead of their regular genes
Avian Myeloblastosis
Virus

R U5 GAG POL MYB U3 R

Feline Sarcoma Virus (FSV)

R U5 dGAG FMS dENV U3 R

Avian Myelocytoma Virus (MC29)

44
R U5 dGAG MYC dENV U3 R
RNA Tumor Viruses

Viral Oncogene
V-onc

Cellular Proto-oncogene
C-onc

45
 RNA Tumor Viruses
Proto-oncogene
A cellular (host) gene that is homologous with a
similar gene that is found in a transforming virus
A cellular oncogene can only induce
transformation after
• mutation
• some other change in the cell’s genome 46
 RNA Tumor Viruses
The discovery of the acutely transforming
retroviruses that contain
v-oncs explains how cancers may arise as a result
of infection

These viruses cause rapid cancer in animals in

the laboratory

47
 RNA Tumor Viruses
In contrast:
Chronically transforming retroviruses
cause tumors inefficiently after prolonged period of time
Avian Leukosis Virus (causes lymphomas)

R U5 GAG POL ENV U3 R

No oncogene! – How does it cause a

tumor? 48
 RNA Tumor Viruses
ALV can integrate into the host cell genome at
MANY locations
but in tumor it is always at the SAME site (or
restricted number of sites)
Suggests tumor arose from one cell
• Something must be important about this site for
transformation
• Crucial event must be rare
49
 RNA Tumor Viruses
What is special about this site?
Myelocytoma tumors from several birds all have
the oncogene close to this site

It is close to
C-myc!
Oncogenesis by promotor insertion
50
 RNA Tumor Viruses

Could C-oncs be involved in NON-VIRAL cancers?

51
 RNA Tumor Viruses
What do oncogenes encode?
Proteins that are involved in growth control and
differentiation

Growth factors
Growth factor receptors
Signal transduction proteins
Transcription factors

52
 DNA Tumor Viruses
How they tumors depended on
our knowledge of RNA tumor
viruses

53
 DNA Tumor Viruses
Herpes
Genes can be
assigned to
sites on
specific
myb mos
chromosomes
myc

mos and myc :

chromosome 8

fes fes: chromosome 15

54
 Cancers often result from gene
Burkitt’s Lymphoma
translocations
8:14 translocation
Break in chromosome
14 at q32

myc

Acute myelocytic leukemia

7:15
9:18 55
11:15:17
 Oncogenesis by rearrangement

Tumor c-onc new promotor

Burkitt’s lymphoma myc (8) Ig heavy (8 to 14)
Ig light (8 to 2)
B-cell chronic lymphocytic bcl-1 Ig heavy (11 to 14)
leukemia bcl-2 Ig heavy (18 to 14)

T cell chronic lymphocytic tcl-1 T cell receptor

leukemia (14 inversion)
T cell chronic lymphocytic myc T cell receptor (8 to 14)
leukemia 56
 Oncogenes
Mutations in a proto-oncogene are dominant “gain
of function” mutations
However other oncogenic genes show recessive
mutations
Anti-Oncogenes
• Loss of function mutations
• Retinoblastoma
• p53 57
 Proto-oncogenes
Dominant
Heterozygote mutations Homozygote

Allele 1 Allele 2 Allele 1 Allele 2

Normal Mutant Mutant Mutant

Binds under Mutant Mutant Mutant

special always always always
circumstances binds binds binds

Always binds Always binds

58
Function gained Function gained
 Anti-Oncogenes
Recessive mutations
Mutation growth

Rb Gene Mutant Rb Mutant Rb Mutant Rb

Rb
Rb protein

Heterozygote Homozygote

Rb Function lost

Binds and controls cell cycle No binding - Growth continues

59
Turns off DNA replication
 Anti-Oncogenes
Retinoblastoma gene has normal
regulatory function in many cells

Involved in
Retinoblastoma
Lung carcinomas
Breast carcinomas 60
Anti-Oncogenes
P53
Inactivated by
• deletion
• point mutation

61
 DNA Tumor Viruses
Oncogenes
• Adenovirus E1A region 2
• SV 40 Large T
• Polyoma Large T
• BK virus Large T
• Lymphotropic virus Large T
• Human papilloma Virus-16 E6, E7

All have a sequence in common

62
Mutations in this region abolish transformation capacity
 Anti-Oncogenes
Retinoblastoma
Rb Gene Adenovirus E1A

Rb Rb
protein 105kD

Rb

Rb
63
Stops replication Cell cycle continues
 Anti-Oncogenes
p53
P53 gene P53 gene P53 gene
Hepatitis C Papilloma

P53 P53 P53

Papilloma
proteolysis
P53 DNA

64
Stops replication replication replication