1 13 Molecular Genetics of Cell Cycle and Cancer

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1 13 Molecular Genetics of Cell Cycle and Cancer Slide 2 2 There are two major parts in the cell cycle: Interphase: G 1 = gap1S = DNA synthesis G 2 = gap2 Mitosis: M There are two essential functions of the cell cycle: To ensure that each chromosomal DNA molecule is replicated only once per cycle To ensure that the identical replicas of each chromosome are distributed equally to the two daughter cells The Cell Cycle Slide 3 3 Fig. 13.1 Slide 4 4 The Cell Cycle The cell cycle is under genetic control A fundamental feature of the cell cycle is that it is a true cycle: it is not reversible Many genes are transcribed during the cell cycle just before their products are needed Mutations affecting the cell cycle have helped to identified the key regulatory pathways Slide 5 5 The Cell Cycle Progression from one phase to the next is propelled by characteristic protein complexes, which are composed of Cyclins and cyclin-dependent protein kinases (CDK) Expression of mitotic cyclins E, A, and B are periodic, whereas cyclin D is expressed throughout the cell cycle in response to mitosis stimulating drugs (mitogens) The cyclin-CDK complexes phosphorylate targeted proteins, changing dramatically their activity Slide 6 6 Fig. 13.8 Slide 7 7 The Retinoblastoma Protein The retinoblastoma (RB) protein controls the initiation of DNA synthesis. RB maintains cells at a point in G1 called the G1 restriction point or start by binding to the transcription factor E2F, until the cell has attained proper size If the cycling cell is growing properly and becomes committed to DNA synthesis, several cyclin-CDK complexes inactivate RB by phosphorylation After cell enters S phase E2F becomes phosphorylated as well and loses its ability to bind DNA Slide 8 8 Fig. 13.10 Slide 9 9 The Cell Cycle The progression from G2 to M is controlled by a cyclin B-CDK2 complex know as maturation- promoting complex Protein degradation (proteolysis) also helps regulate the cell cycle. The anaphase-promoting complex (APC/C), which is a ubiquitinprotein ligase responsible for adding the 76-amino-acid protein ubiquitin to its target proteins and marking them for destruction in the proteasome Proteolysis eliminates proteins used in the preceding phase as well as proteins that would inhibit progression into the next Slide 10 10 Checkpoints Cells monitor their external environments and internal state and functions Checkpoints in the cell cycle serve to maintain the correct order of steps as the cycle progresses; they do this by causing the cell cycle to pause while defects are corrected or repaired Checkpoints in the cell cycle allow damaged cells to repair themselves or to self-destruct Slide 11 11 Three Main Checkpoints A DNA damage checkpoint A centrosome duplication checkpoint A spindle checkpoint Slide 12 12 Fig. 13.12 Slide 13 13 A DNA Damage Checkpoint A DNA damage checkpoint arrests the cell cycle when DNA is damaged or replication is not completed. In animal cells, a DNA damage checkpoint acts at three stages in the cell cycle: at the G1/S transition, in the S period and at the G2/M boundary The p53 transcription factor is a key player in the DNA damage checkpoint. Slide 14 14 A DNA Damage Checkpoint In normal cells, level of activated p53 is very low Protein Mdm2 keeps p53 inactivated by preventing phosphorylation and acetylation of p53 and by exporting p53 from the nucleus Damaged DNA leads to activation of p53 and its release from Mdm2 Slide 15 15 A DNA Damage Checkpoint Activated p53 triggers transcription of a number of genes - p21, 14-3-3s, Bax, Apaf1, Maspin,GADD45 DNA damage detected in G1 blocks cell G1/S transition DNA damage in S phase reduces processivity of DNA polymerase and gives the cell time for repair Processivity: number of consecutive nucleotides that replicate before polymerase detaches from template DNA damage detected in S or G1 arrests cells at G2/M transition Slide 16 16 Fig. 13.15 Slide 17 17 A DNA Damage Checkpoint DNA damage also triggers activation of a pathway for apoptosis = programmed cell death When the apoptotic pathway is activated, a cascade of proteolysis is initiated that culminates in cell suicide The proteases involved are called caspases Slide 18 18 Centrosome Duplication Checkpoint Monitors spindle formation Functions to maintain the normal complement of chromosomes Sometimes coordinates with the spindle checkpoint and the exit from mitosis Slide 19 19 The Spindle Checkpoint Monitors assembly of the spindle and its attachment to kinetochores The kinetochore is the spindle-fiber attachment site on the chromosome Incorrect or unbalanced attachment to the spindle activates spindle checkpoint proteins, triggers a block in the separation of the sister chromatids by preventing activation of the anaphase-promoting complex (APC/C) Slide 20 20 Fig. 13.18 Slide 21 21 Cancer Cancer cells have a small number of mutations that prevent normal checkpoint function Cancer is not one disease but rather many diseases with similar cellular attributes All cancer cells show uncontrolled growth as a result of mutations in a relatively small number of genes Cancer is a disease of somatic cells Slide 22 22 1% of cancer cases are familial: show evidence for segregation of a gene in pedigree 99% are sporadic: the result of genetic changes in somatic cells Within an organism, tumor cells are clonal, which means that they are descendants from a single ancestral cell that became cancerous. Cancer Slide 23 23 Cancer Cells vs. Normal Cells In normal cells, cell-to-cell contact inhibits further growth and division, a process called contact inhibition Cancer cells have lost contact inhibition: they continue to grow and divide, and they even pile on top of one another Slide 24 24 Cancer Cells vs. Normal Cells Even in the absence of damage, normal cells cease to divide in culture after about 50 doublings = cell senescence Senescence of normal cells is associated with a loss of telomerase activity: the telomeres are no longer elongated, which contributes to the onset of senescence and cell death Cancer cells have high levels of telomerase, which help to protect them from senescence, making them immortal Slide 25 25 Key Mutational Targets Many cancers are the result of alterations in cell cycle control, particularly in control of the G1-to-S transition These alterations also affect apoptosis through their interactions with p53 The major mutational targets for the multistep cancer progression are of two types: Proto-oncogenes Tumor-suppressor genes Slide 26 26 Key Mutational Targets The normal function of proto-oncogenes is to promote cell division or to prevent apoptosis The normal function of tumor-suppressor genes is to prevent cell division or to promote apoptosis Slide 27 27 Oncogenes Oncogenes are derived from normal cellular genes called proto-oncogenes Oncogenes are gain-of-function mutations associated with cancer progression Oncogenes are gain-of-function mutations because they improperly enhance the expression of genes that promote cell proliferation or inhibit apoptosis Slide 28 28 Mdm2: Amplification of Mdm2 gene and overexpression of Mdm2 protein leads to inactivation of p53 gene Amplification of Mdm2 gene has been found in many tumors of adipose tissue, soft tissue sarcomas, osteosarcoma, and esophageal carcinoma Cyclin D and CDK4: Amplification and overexpression leads to unscheduled entry to S phase Amplification and/or overexpression has been found in many esophageal carcinomas, bladder and breast cancers Oncogenes Slide 29 29 Oncogenes Growth-factor receptors: Cellular growth factors stimulate growth by binding to a growth-factor receptor at the cell membrane The binding activates a signal transduction pathway that acts through Ras, cyclin D, and its partner CDKs. Amplification and overexpression of the gene that encodes the receptor for epidermal growth factor (EGFR) has been found in many malignant astrocytomas, glioblastomas, breast and ovarian cancers, head and neck cancers and melanomas. Slide 30 30 Oncogenes Ras: The Ras protein acts as a switch in stimulating cellular growth in the presence of growth factors Certain mutant Ras proteins lack GTPase activity and remain in the form of RasGTP. The signal for cellular growth is transmitted constitutively--unrestrained growth and division take place Fig. 13.18 Slide 31 31 Tumor-Suppressor Genes Tumor-suppressor genes normally negatively control cell proliferation or activate the apoptotic pathway Loss-of-function mutations in tumor-suppressor genes contribute to cancer progression. Slide 32 32 p53: Loss of function of p53 eliminates the DNA checkpoint that monitors DNA damage in G1 and S The damaged cells survive and proliferate and their genetic instability increases the probability of additional genetic changes and thus progression toward the cancerous state p53 proves to be nonfunctional in more than half of all cancers Mutant p53 proteins are found frequently in melanomas, lung cancers, colorectal tumors, bladder and prostate cancers, and astrocytomas. Tumor-Suppressor Genes Slide 33 33 p21: Loss of p21 function results in renewed rounds of DNA synthesis without mitosis, and the level of ploidy of the cell increases. Mutations in the p21 gene occur in some prostate cancers. p16/p19ARF: The p16 and p19ARF proteins are products of the same gene transcribed from different promoters The p16 product can inhibit the cyclin DCdk4 complex and help control entry into S phase The gene is deleted in many gliomas, mesotheliomas, melanomas, nasopharyngeal carcinomas, biliary-tract and esophageal carcinomas. Tumor-Suppressor Genes Slide 34 34 RB: The retinoblastoma protein controls the transition from G1 to S phase by controlling the activity of the transcription factor E2F Loss of RB function frees E2F hence excessive rounds of DNA synthesis are continuously being initiated. Loss of RB function is found in melanomas, small-cell lu