Tumors and cancer: Section outline

• Mammalian tumors
• Basic mechanisms that lead to tumor formation
• Cancer critical genes (oncogenes and tumor suppressor genes)
• Cancer genomics
• Comparison between mammalian and plant tumors
• Crown gall tumors in plants and Agrobacterium tumefaciens
• The discovery of interkingdom lateral gene transfer
• Plant oncogenes of bacterial origin
• Molecular mechanism of lateral gene transfer
• Exploitation of the system for generating transgenic plants
• Examples of transgenic plants used for food production
 Plant tumors and transgenic plants

• Why studying plant tumors on the molecular level?

• Understanding the mechanisms
underlying cellular differentiation
• Best studied example of
inter-kingdom lateral gene transfer
• Led to the primary tool for generating
genetically modified plants
 Tumors and cancer

• The phenotype:
• Tumor: An abnormal mass of tissue that results from excessive cell division or
reduced cell death. Can be benign or malignant
• Benign: No invasion to nearby tissue or spreading to other parts of the body
• Malignant: Tendency to invade and destroy nearby tissue and/or spread
to other parts of the body (forms metastases)
• Cancer: A malignant tumor

Merkel cell tumor Crown gall tumor

(on human): a rare (on elm tree): a
malignant tumor of the common tumor of
skin stems and roots
 Animal cancer

Alberts (2015) Molecular Biology of the

cell. 6th edition, Figure 20-1 and 20-13
• cancer cells reproduce without any normal restraints on cell
growth, cell division, or programmed cell death
• cancer cells invade tissues normally belonging to other cells
types (eventually forming metastasis)
• cancers evolve gradually
• contain increasing numbers of somatic
mutations and epigenetic alterations
• cause malfunctioning of regulatory
proteins leading to
• increased cell division
• decreased cell death
 Tumor progression
• Slow progression:
• exposure to mutagens / carcinogens increases risk
to develop cancer
• years pass by from initial exposure to formation
of cancer
• tumor progression:
• normal tissue (A)

Alberts (2015) Molecular Biology of the

cell. 6th edition, Figure 20-7 and 20-8
• mild disorder (B)
• benign tumor (not invasive, C)
• malign cancer (metastases, D)
à a single mutation in one gene is
not enough to cause cancer
 Animal cancer and cell cycle control

• mutated genes play key roles in:

• DNA repair
• cell signalling
• cell growth and differentiation
• cell cycle control
• G1-to S checkpoint
• is environment favourable?
• G2-to-M checkpoint
• is all DNA replicated
• is environment favourable
Alberts (2015) Molecular Biology
of the cell. 6th edition, Figure 17-4
 Genetic instability

• Pre-cancer cells accumulate more mutations than normal

cells
• initial mutations may interfere with:
• DNA repair
• DNA damage response (cell cycle arrest)
• epigenetic control mechanisms
• heterochromatine organization

Alberts (2015) Molecular Biology of the

• DNA methylation

cell. 6th edition, Figure 20-10

à increased mutation rate (cells are more likely to
accumulate additional mutations / epigenetic changes
à cells become genetically unstable
Subsequent mutations – cancer critical genes
• additional mutations required that cause cells to:
• constitutively activate cell division in absence of growth factors
• become insensitive to external and internal signals: Alberts (2015) Molecular Biology of the
cell. 6th edition, Figure 20-15

• anti-proliferating factors
• apoptosis inducing factors
• factors inducing differentiation
• factors preventing proliferation without adhesion
• induce ‘help’ from normal tissues
• supporting cells produce growth factors
and remodel extracellular matrix
• induction of angiogenesis
à become invasive (survive and proliferate in other tissues)
Alberts (2015) Molecular Biology of the
Metastasis

cell. 6th edition, Figure 20-16

What Do We Have So Far?

• mutations, mutations, mutations, and epigenetic alterations!

à genetic instability
à more mutations
• mutations allow cells to overcome regulatory
mechanism that regulate
• cell division
• apoptosis
• differentiation
(allows cell to thrive in ‘foreign’ environments)
 Cancer critical genes

• genes that - when altered -

contribute to cancer development:
• gain of function: oncogenes
(dominant, overactive /
overexpressed)
• loss of functions: tumor-
suppressor genes (recessive,
underactive/missing)

Alberts (2015) Molecular Biology of the

cell. 6th edition, Figure 20-17
 Proto-oncogene to oncogene mutations
• possible mechanisms that over-activate a gene:
• mutation in coding region
• mutation in promoter/enhancer
• gene amplification
• chromosomal rearrangements
loss of regulation on protein level
(post-translational modifications)
miss-regulated expression
(over-expression)
of the cell. 6th edition, Figure 20-18
Alberts (2015) Molecular Biology
 murine leukemia virus
Oncogenes vectored by Retroviruses

• oncogenes can be introduced by retroviruses:

Rous sarcoma virus (RSV, causes
• retrovirus integrates into genomic DNA connective-tissue tumors: sarcomas)
• maybe close to a (proto-) oncogene
• when excised it may take (part) of the proto-
oncogene with it contains in addition a Tyrosin kinase (v-Src),
which is responsible for cancer formation
• may mutate to oncogene while in
retrovirus • v-SRC is a mutated version of c-SRC
(found in the host)
• retrovirus integrates itself plus oncogene into • c-SRC is a protein kinase involved in
new host regulating embryo development / cell
proliferation
• oncogene contributes to cancer formation
• extremely unlikely to happen, but once it does it
is a great selective advantage for retrovirus...
Alberts (2008) Molecular Biology
of the cell. 5th edition, Figure 20-28
• possible mechanisms that lead to loss of function of a gene: Loss of function
• first allele: mutations of tumor-
• mutation in coding region protein non-functional suppressor genes
• deletion
• mutation in promoter/enhancer silenced first allele normal second allele
• epigenetic silencing
• second allele:

Biology of the cell. 5th edition, Figure 20-21

modified from Alberts (2008) Molecular
 Loss of function mutations of tumor-suppressor genes
• mutated versions of tumor-suppressor genes may be inherited
• increased likelihood of developing cancer
• rare hereditary form of cancer used to identify tumor suppressor genes:
• example:
Retinoblastoma [Rb]:

modified from Alberts (2008) Molecular

Biology of the cell. 5th edition, Figure 20-20
 Rb is a suppressor of E2F

• E2F:
• Rb encodes a protein with protein-
protein interaction domain • family of transcription factors
• interacts with E2F • activate transcription of many
S-phase related gene
• E2F inactive when bound to Rb
• replication origin binding
• Rb/E2F inhibits S-phase gene
proteins, DNA polymerase,
expression
nucleotide biosynthetic
Rb is a suppressor of E2F genes, cell cycle regulatory
Rb is the block preventing genes
cells from entering S-phase
E2F is a major switch
activating S-phase
 Rb mediated activation of S-phase

• mitogen binding activates a signalling loss of Rb function

cascade
constitutively active E2F
• leads to the activation of specific
protein kinases (called S-phase Cyclin loss of G1-S checkpoint
dependent Kinase, S-CdK)
• S-CdK phosphorylates Rb
• phosphorylated Rb does no longer
bind E2F
• E2F activates transcription of S-phase
specific genes
à cell enters S-phase

modified from Alberts (2015) Molecular

Biology of the cell. 6th edition, Figure 17-61
 Ras: a virus transmitted oncogene
• also involved in cell cycle control (entry into S-phase)
• encodes a small GTP binding protein / GTPase
• member of large gene family
• ubiquitous signal transduction component
• activated by guanine nucleotide
exchange factors (GEFs)
• lead to the activation of downstream
signalling components (e.g. MAP kinase)

of the cell. 6th edition, Figure 15-8

Alberts (2015) Molecular Biology
• inactivated by GTPase activating enzymes (GAPs)
• GAPs enhance intrinsic GTPase activity
 MAP kinases: also oncogenes

of the cell. 6th edition, Figure 15-7

Alberts (2015) Molecular Biology
Ras (inactive)

• mitogen activates Ras

• Ras initiates Mitogen Activated
Protein kinase cascade Ras (active)
• Ras activates MAP kinase kinase kinase MAP kinase (inactive)

à phosphorylates MAP kinase kinase

à phosphorylates MAP kinase
MAP kinase
à phosphorylates target proteins kinase (active)
(e.g. transcription factors)

MAP kinase
(active)
 S-phase activation pathway

Alberts (2015) Molecular Biology of

the cell. 6th edition, Figure 17-61
• binding of a mitogen to a cell surface receptor leads to
activation of a specific GEF
• leads to activation of Ras
• activated Ras initiates a MAPK cascade
• activated MAPK phosphorylates transcription factor
• expression of early genes (including another
transcription factor: Myc)
• MYC activates transcription of delayed response genes
(including G1/S cyclin)
• cyclin binds to a cyclin dependent-kinase (CdK)
• G1/Cdk complex phosphorylates Rb
• releases E2F à activation of S-phase gene
transcription
 The p53 tumor suppressor gene
• not essential for development
• homozygous NULL mutants (mouse) appear normal
• EXCEPTION: develop cancer early (before 10 month of age)
• p53 protein amounts increase in response to stress (UV, g-rays)
Alberts (2015) Molecular Biology of
the cell. 6th edition, Figure 20-27

• p53 is a transcription factor

• most cancerous loss-of
function mutations are in
DNA binding domain
 The p53 transcriptional activator (non-mutated)

• DNA damage induces (somehow) a kinase cascade

• p53 is phosphorylated and thereby activated

• one target of p53 transcriptional

activation is a protein called p21
• p21 binds to the G1/S-phase
Alberts (2015) Molecular Biology of

activated Cyclin dependent

the cell. 6th edition, Figure 17-62

kinase (G1/S-CdK), inactivating it

p53 mediated inhibition

Alberts (2015) Molecular Biology of

the cell. 6th edition, Figure 17-61
of S-phase activation
pathway

• a primary function of the p53 pathway is

preventing damaged cells to enter S-phase
à p53 is a true tumor suppressor gene
 There are many more proto-oncogenes and tumor suppressor genes

• we just took a
glimpse into the
signal-transduction
network regulating
cell-proliferation
and cell death…
 Summary: Animal Cancer

• caused by somatic mutations and epigenetic changes

• cause genetic instability
• interfere with regulatory network suppressing or activating cell
proliferation and cell death
• multiple mutations are necessary
• overcome built in protective mechanisms
• leads to excessive cell-proliferation (tumor)
• additional mutations necessary for a tumor to become invasive and
to form metastases (cancer)
• most cancer critical genes are involved in signal transduction mediating
control of cell cycle, differentiation, and cell death