1. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings PowerPoint Lectures for Biology, Seventh Edition Neil Campbell and Jane Reece Lectures by Chris Romero Chapter 12 & 13 The Cell Cycle

2. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Overview: The Key Roles of Cell Division The continuity of life – Is based upon the reproduction of cells, or cell division Figure 12.1

3. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Unicellular organisms often reproduce by mitotic cell division (asexual) …but so do others 100 µm An amoeba, a single-celled eukaryote, is dividing into two cells. Each new cell will be an individual organism. Spores budding Vegetative propagation Animation Animation of bacterial DNA replication = Binary fission.

4. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Multicellular organisms depend on cell division for – Development from a fertilized cell – Growth – Repair 20 µm200 µm (b) Growth and development. This micrograph shows a sand dollar embryo shortly after the fertilized egg divided, forming two cells (LM). (c) Tissue renewal. These dividing bone marrow cells (arrow) will give rise to new blood cells (LM). Figure 12.2 B, C

5. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings CELL DIVISION MITOTIC cell division results in genetically identical daughter cells Cells duplicate their genetic material – Before they divide, ensuring that each daughter cell receives an exact copy of the genetic material, DNA – Is an integral part of the cell cycle

6. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cellular Organization of the Genetic Material A cell’s endowment of DNA, its genetic information – Is called its genome 50 µm The DNA molecules in a cell – Are packaged into chromosomes

7. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Eukaryotic chromosomes Consist of chromatin, a complex of DNA and protein that condenses during cell division In animals – Somatic cells have two sets of chromosomes Diploid (2n) – Gametes have one set of chromosomes Haploid (n)

8. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Distribution of Chromosomes During Cell Division Each duplicated chromosome h as two sister chromatids, which separate during cell division 0.5 µm Chromosome duplication (including DNA synthesis) Centromere Separation of sister chromatids Sister chromatids Centromeres Sister chromatids A eukaryotic cell has multiple chromosomes, one of which is represented here. Before duplication, each chromosome has a single DNA molecule. Once duplicated, a chromosome consists of two sister chromatids connected at the centromere. Each chromatid contains a copy of the DNA molecule. Mechanical processes separate the sister chromatids into two chromosomes and distribute them to two daughter cells. Figure 12.4 In preparation for cell division – DNA is replicated and the chromosomes condense

9. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Eukaryotic cell division consists of Mitosis, the division of the nucleus – Cytokinesis, the division of the cytoplasm In meiosis – Sex cells are produced after a reduction in chromosome number

10. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Phases of the Cell Cycle The cell cycle consists of – The mitotic phase – Interphase INTERPHASE G1G1 S (DNA synthesis) G2G2 Cytokinesis Mitosis MITOTIC (M) PHASE Figure 12.5 Notice the time spent in interphase

11. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings What happens during Interphase? Interphase can be divided into subphases – G 1 phase – S phase – G 2 phase The G 0 phase (referred to the G zero phase) or resting phase is a period in the cell cycle in which cells exist in a quiescent state. G 0 phase is viewed as either an extended G 1 phase, where the cell is neither dividing nor preparing to divide, or a distinct quiescent stage that occurs outside of the cell cycle. Some types of cells, such as nerve and heart muscle cells, become quiescent when they reach maturity.

12. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings MITOSIS The mitotic phase – Is made up of mitosis and cytokinesis

13. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings PMAT Mitosis consists of four distinct phases – Prophase G 2 OF INTERPHASE PROPHASE PROMETAPHASE Centrosomes (with centriole pairs) Chromatin (duplicated) Early mitotic spindle Aster Centromere Fragments of nuclear envelope Kinetochore Nucleolus Nuclear envelope Plasma membrane Chromosome, consisting of two sister chromatids Kinetochore microtubule Figure 12.6 Nonkinetochore microtubules

14. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings – Metaphase – Anaphase – Telophase Centrosome at one spindle pole Daughter chromosomes METAPHASEANAPHASETELOPHASE AND CYTOKINESIS Spindle Metaphase plate Nucleolus forming Cleavage furrow Nuclear envelope forming Figure 12.6

15. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Mitotic Spindle: A Closer Look The mitotic spindle – Is an apparatus of microtubules that controls chromosome movement during mitosis The spindle arises from the centrosomes – And includes spindle microtubules and asters

16. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Some spindle microtubules – Attach to the kinetochores of chromosomes and move the chromosomes to the metaphase plate CentrosomeAster Sister chromatids Metaphase Plate Kinetochores Overlapping nonkinetochore microtubules Kinetochores microtubules Centrosome Chromosomes Microtubules 0.5 µm 1 µm Figure 12.7

17. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings In anaphase, sister chromatids separate – And move along the kinetochore microtubules toward opposite ends of the cell EXPERIMENT 1 The microtubules of a cell in early anaphase were labeled with a fluorescent dye that glows in the microscope (yellow). Spindle pole Kinetochore Figure 12.8

18. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Nonkinetechore microtubules from opposite poles – Overlap and push against each other, elongating the cell In telophase – Genetically identical daughter nuclei form at opposite ends of the cell

19. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cytokinesis: A Closer Look In animal cells – Cytokinesis occurs by a process known as cleavage, forming a cleavage furrow Cleavage furrow Contractile ring of microfilaments Daughter cells 100 µm (a) Cleavage of an animal cell (SEM) Figure 12.9 A

20. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings In plant cells, during cytokinesis – A cell plate forms Daughter cells 1 µm Vesicles forming cell plate Wall of patent cell Cell plate New cell wall (b) Cell plate formation in a plant cell (SEM) Figure 12.9 B

21. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Mitosis in a plant cell 1 Prophase. The chromatin is condensing. The nucleolus is beginning to disappear. Although not yet visible in the micrograph, the mitotic spindle is staring to from. Prometaphase. We now see discrete chromosomes; each consists of two identical sister chromatids. Later in prometaphase, the nuclear envelop will fragment. Metaphase. The spindle is complete, and the chromosomes, attached to microtubules at their kinetochores, are all at the metaphase plate. Anaphase. The chromatids of each chromosome have separated, and the daughter chromosomes are moving to the ends of cell as their kinetochore microtubles shorten. Telophase. Daughter nuclei are forming. Meanwhile, cytokinesis has started: The cell plate, which will divided the cytoplasm in two, is growing toward the perimeter of the parent cell. 23 4 5 Nucleus Nucleolus Chromosome Chromatine condensing Figure 12.10

22. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Mitosis and cytokinesis narrated animationMitosis and cytokinesis How the Cell Cycle Works narrated animationHow the Cell Cycle Works

23. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Cell Cycle Control System The sequential events of the cell cycle – Are directed by a distinct cell cycle control system, which is similar to a clock Figure 12.14 Control system G 2 checkpoint M checkpoint G 1 checkpoint G1G1 S G2G2 M

24. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The clock has specific checkpoints – Where the cell cycle stops until a go-ahead signal is received G 1 checkpoint G1G1 G1G1 G0G0 (a) If a cell receives a go-ahead signal at the G 1 checkpoint, the cell continues on in the cell cycle. (b) If a cell does not receive a go-ahead signal at the G 1 checkpoint, the cell exits the cell cycle and goes into G 0, a nondividing state. Figure 12.15 A, B

25. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Regulation of the Cell Cycle The frequency of cell division – Varies with the type of cell – The cell cycle is regulated by a molecular control system Molecules present in the cytoplasm – Regulate progress through the cell cycle Two types of regulatory proteins are involved in cell cycle control Cyclins Cyclin-dependent kinases (Cdks) Narrated animation “Control of the Cell Cycle”Control of the Cell Cycle

26. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cell division is tightly controlled by complexes made of several specific proteins. These complexes contain enzymes called cyclin- dependent kinases (CDKs), which turn on or off the various processes that take place in cell division. CDK partners with a family of proteins called cyclins. One such complex is mitosis-promoting factor (MPF), sometimes called maturation-promoting factor, which contains cyclin A or B and cyclin- dependent kinase (CDK). (See Figure 2a.) CDK is activated when it is bound to cyclin, interacting with various other proteins that, in this case, allow the cell to proceed from G2 into mitosis. The levels of cyclin change during the cell cycle (Figure 2b). In most cases, cytokinesis follows mitosis. Narrated animation: Cell Proliferation Signaling Pathway http://highered.mcgraw- hill.com/olc/dl/120073/bio15.swfhttp://highered.mcgraw- hill.com/olc/dl/120073/bio15.swf

27. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings As shown in Figure 3, different CDKs are produced during the phases. The cyclins determine which processes in cell division are turned on or off and in what order by CDK. As each cyclin is turned on or off, CDK causes the cell to move through the stages in the cell cycle.

28. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Both internal and external signals Control the cell cycle checkpoints There are three checkpoints a cell must pass through: the G1 checkpoint, G2 checkpoint, and the M-spindle checkpoint (Figure 4). At each of the checkpoints, the cell checks that it has completed all of the tasks needed and is ready to proceed to the next step in its cycle. Cells pass the G1 checkpoint when they are stimulated by appropriate external growth factors; for example, platelet-derived growth factor (PDGF) stimulates cells near a wound to divide so that they can repair the injury. The G2 checkpoint checks for damage after DNA is replicated, and if there is damage, it prevents the cell from going into mitosis. The M-spindle (metaphase) checkpoint assures that the mitotic spindles or microtubules are properly attached to the kinetochores (anchor sites on the chromosomes). If the spindles are not anchored properly, the cell does not continue on through mitosis. The cell cycle is regulated very precisely. Mutations in cell cycle genes that interfere with proper cell cycle control are found very often in cancer cells.

29. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Growth Factor proteins Growth factors – Stimulate other cells to divide EXPERIMENT A sample of connective tissue was cut up into small pieces. Enzymes were used to digest the extracellular matrix, resulting in a suspension of free fibroblast cells. Cells were transferred to sterile culture vessels containing a basic growth medium consisting of glucose, amino acids, salts, and antibiotics (as a precaution against bacterial growth). PDGF was added to half the vessels. The culture vessels were incubated at 37°C. 3 2 1 Petri plate Without PDGF With PDGF Scalpels Figure 12.17

30. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Environmental effects on cell cycling In density-dependent inhibition – Crowded cells stop dividing Most animal cells exhibit anchorage dependence – In which they must be attached to a substratum to divide Cells anchor to dish surface and divide (anchorage dependence). When cells have formed a complete single layer, they stop dividing (density-dependent inhibition). If some cells are scraped away, the remaining cells divide to fill the gap and then stop (density-dependent inhibition). Normal mammalian cells. The availability of nutrients, growth factors, and a substratum for attachment limits cell density to a single layer. (a) 25 µm Figure 12.18 A

31. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cancer cells – Exhibit neither density-dependent inhibition nor anchorage dependence 25 µm Cancer cells do not exhibit anchorage dependence or density-dependent inhibition. Cancer cells. Cancer cells usually continue to divide well beyond a single layer, forming a clump of overlapping cells. (b) Most oncogenes are mutations of certain normal genes called proto-oncogenes. Proto-oncogenes are the "good" genes that normally control what kind of cell it is and how often it divides. When a proto-oncogene mutates (changes) into an oncogene, it becomes a "bad" gene that can become permanently turned on or activated when it is not supposed to be. When this happens, the cell grows out of control, which can lead to cancer.

32. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings How cell division (and thus tissue growth) is controlled is very complex. The following terms are some of the features that are important in regulation, and places where errors can lead to cancer. Cancer is a disease where regulation of the cell cycle goes awry and normal cell growth and behavior is lost. Cdk (cyclin dependent kinase, adds phosphate to a protein), along with cyclins, are major control switches for the cell cycle, causing the cell to move from G1 to S or G2 to M. MPF (Mitosis Promoting Factor) includes the CdK and cyclins that triggers progression through the cell cycle. p53 is a protein that functions to block the cell cycle if the DNA is damaged. If the damage is severe this protein can cause apoptosis (cell death). p53 levels are increased in damaged cells. This allows time to repair DNA by blocking the cell cycle. A p53 mutation is the most frequent mutation leading to cancer. An extreme case of this is Li Fraumeni syndrome, where a genetic a defect in p53 leads to a high frequency of cancer in affected individuals. p27 is a protein that binds to cyclin and Cdks, blocking entry into S phase. Recent research (Nature Medicine 3, 152 (1997)) suggests that breast cancer prognosis is determined by p27 levels. Reduced levels of p27 predict a poor outcome for breast cancer patients. Loss of cell cycle control

33. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Loss of Cell Cycle Controls in Cancer Cells Cancer cells – Do not respond normally to the body’s control mechanisms – Form tumors P53 is a tumor suppressor protein that in humans is encoded by the TP53 gene Ras proteins also play a role in cell growth and division. Overactive Ras signaling can ultimately lead to cancer ( ra t s arcoma) BRCA1 and BRCA 2 are tumor-suppressor genes expressed in the cells of breast and other tissue, where it helps repair damaged DNA, or destroy cells if DNA cannot be repaired. If BRCA itself is damaged, damaged DNA is not repaired properly and this increases risks for cancers

34. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Malignant tumors invade surrounding tissues and can metastasize – Exporting cancer cells to other parts of the body where they may form secondary tumors Cancer cells invade neighboring tissue. 2 A small percentage of cancer cells may survive and establish a new tumor in another part of the body. 4 Cancer cells spread through lymph and blood vessels to other parts of the body. 3 A tumor grows from a single cancer cell. 1 Tumor Glandular tissue Cancer cell Blood vessel Lymph vessel Metastatic Tumor Figure 12.19

35. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Case of Henrietta Lacks HeLa cells and their legacy 1920–1951 Who was she? Familiarize yourself with her biography. What was her significance to medical science? What were the ethical questions her case raises?

36. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Meiosis Overview Concept 13.3: Meiosis reduces the number of chromosome sets from diploid to haploid – This maintains the correct # of chromosomes in each organism of a species Meiosis introduces genetic variation into offspring (without changing the chromosome #)

37. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Meiosis leads to greater genetic variation in offspring Concept 13.4: Genetic variation produced in sexual life cycles contributes to evolution Reshuffling of genetic material during synapsis of Meiosis I (crossing over) animationanimation Independent assortment (animation) of homologues and chromatids ensures that gametes will all be uniqueanimation – Produces genetic variation How does this ensure natural selection can take place? Examples?

38. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Overview Meiosis takes place in two sets of divisions Meiosis I – Replicates the diploid number of chromosomes and recombines them randomly Meiosis II – Reduces the number of chromosomes from diploid to haploid – Narrated animation of the stages of meiosis stages of meiosis

39. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Centrosomes (with centriole pairs) Sister chromatids Chiasmata Spindle Tetrad Nuclear envelope Chromatin Centromere (with kinetochore) Microtubule attached to kinetochore Tertads line up Metaphase plate Homologous chromosomes separate Sister chromatids remain attached Pairs of homologous chromosomes split up Chromosomes duplicate Homologous chromosomes (red and blue) pair and exchange segments; 2n = 6 in this example INTERPHASE MEIOSIS I: Separates homologous chromosomes PROPHASE I METAPHASE I ANAPHASE I Interphase and meiosis I Figure 13.8

40. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings TELOPHASE I AND CYTOKINESIS PROPHASE II METAPHASE II ANAPHASE II TELOPHASE II AND CYTOKINESIS MEIOSIS II: Separates sister chromatids Cleavage furrow Sister chromatids separate Haploid daughter cells forming During another round of cell division, the sister chromatids finally separate; four haploid daughter cells result, containing single chromosomes Two haploid cells form; chromosomes are still double Figure 13.8 Telophase I, cytokinesis, and meiosis II

41. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Spermatogenesis and Oogenesis

42. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings SUMMARY: Mitosis vs Meiosis Similarities? Differences? Animated comparison between mitosis and meiosisAnimated comparison Figure 13.9 MITOSIS MEIOSIS Prophase Duplicated chromosome (two sister chromatids) Chromosome replication Chromosome replication Parent cell (before chromosome replication) Chiasma (site of crossing over) MEIOSIS I Prophase I Tetrad formed by synapsis of homologous chromosomes Metaphase Chromosomes positioned at the metaphase plate Tetrads positioned at the metaphase plate Metaphase I Anaphase I Telophase I Haploid n = 3 MEIOSIS II Daughter cells of meiosis I Homologues separate during anaphase I; sister chromatids remain together Daughter cells of meiosis II n n nn Sister chromatids separate during anaphase II Anaphase Telophase Sister chromatids separate during anaphase 2n2n2n2n Daughter cells of mitosis 2n = 6

43. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings A Comparison of Mitosis and Meiosis Meiosis and mitosis can be distinguished from mitosis by three events in Meiosis l 1.Synapsis and crossing over – Homologous chromosomes physically connect and exchange genetic information 2.Tetrads on the metaphase plate – At metaphase I of meiosis, paired homologous chromosomes (tetrads) are positioned on the metaphase plates 3.Separation of homologues – At anaphase I of meiosis, homologous pairs move toward opposite poles of the cell – In anaphase II of meiosis, the sister chromatids separate – Flash animation of these events Flash animation of these events