Tumor Suppressor Definition: Proteins that negatively regulate the cell cycle.

Not inclusive definition: DNA repair proteins do not negatively regulate cell cycle

Two Major Tumor Suppressors that we have discussed:

          pRb; inhibits E2F-dependent cell cycle transcription

          p53; activated by DNA damage to enforce cell cycle arrest and/or cell death

In many familial cancer syndromes, affected individuals inherit a single functional

copy of a tumor suppressor gene, and this functional copy is inactivated.

Tumor Suppressors can be inactivated by several mechanisms

          A second mutation

Promoter methylation

Loss of Heterozygosity (LOH)

DNA Tumor Virus Oncoproteins

Centrosomes are Critical for Mitotic Fidelity

          Cells inherit a single centrosome through mitosis and cytokinesis

          this centrosome must be duplicated exactly once to form the mitotic spindle

Centrosome abnormalities are seen in most human tumors

          Centrosome abnormalities cause mitotic abnormalities

High risk Human Papiloma Virus (HPV) inactivates pRb and p53

          Integration of virus (happens only rarely) misregulates viral gene expression

                    only E6 and E7 are expressed

E6 and E7 cooperatively transformation cells

Overexpression of E6 and E7 allow accumulation of DNA damage

          E6 inhibits p53, cells cannot halt cell cycle in response to DNA damage

          E7 inhibits pRb, rapid S-phase progression may actually cause damage

High risk HPV (i.e. HPV16 and HPV18) causes over-duplication of centrosomes

          Extra centrosomes cause mitotic defects:

                    multi-polar spindles, unaligned chromosomes, lagging chromosomes

E7-expressing cells accumulate extra centrosomes before becoming aneuploid

          Inhibition of pRB allows E2F-dependent expression of Cdk2, cyclins E and A

                    all accelerate centrosome duplication

          E7-expressing cells over-duplicate centrosomes within a single cell cycle

E6-expressing cells become aneuploid before accumulating extra centrosomes

          Inhibition of p53 causes cells to fail to arrest in G2 due to DNA damage

                    this leads to frequent failure of cytokinesis, producing polyploidy cells

          Inhibition of p53 allows cells that failed in cytokinesis to enter the cell cycle

                    these cells inherit two nuclei (polypoloidy) and TWO centrosomes

          Inhibition of p53 therefore indirectly causes accumulation of centrosomes

E6 and E7 cooperate to transform cells

          E6 inhibits p53, producing polyploidy cells

(extra copies of all chromosomes and two centrosomes)

          E7 inhibits pRb, allowing overduplication of centrosomes

          Extra Centrosomes Cause Mitotic Abnormalities in Polyploid Cells

                    This causes aneuploidy, which can drive transformation

                    (by unbalancing positive and negative cell cycle regulators)