Concept


In a biological sense, growth results from the reproduction of new cells from pre-existing ones, by the process of cell division (mitosis). Once a tissue or organ reaches an appropriate size, mitosis slows and cells enter a resting phase. This cell cycle of growth and rest is controlled by "checkpoint" molecules first characterized in the 1980s and 1990s in yeast, and then in other eukaryotes. Remarkably, normal development requires that some healthy cells be eliminated, killed, by a process called apoptosis. Initial clues about the nature of apoptosis came from detailed studies of the roundworm Caenorhabditis elegans, in which development of each of the 959 cells in the adult can be traced from the fertilized egg. Analysis of cell "fates" showed that specific cells are programmed to die at specific times during embryonic development. Disruptions in the program leads to an overabundance of cells — a hallmark of cancer.

Animation


Hi, I'm Leland Hartwell. I was one of the first to use yeast cells as a model system to study biological problems. I was interested in how cells regulate the timing of growth and cell division. Yeasts are single-celled organisms that divide by budding. The process is the same as mitosis except that the nuclear membrane stays intact. The yeast cells in this photo express a fluorescent protein in their membranes, and you can see budding as well as non-budding yeast cells. Through my work with yeast cells, I found that all eukaryotic cells cycle through four different stages. A cell first has to grow and replenish its resources. This stage is called G1; the G stands for "gap." Next, the cell synthesizes DNA in preparation for cell division. This is the S stage. After DNA replication, the cell enters the second gap stage, G2, where it makes other proteins and cellular components necessary for cell division. M is the mitotic stage; the cell divides and the whole cycle repeats. Mitosis is dependent on the completion of all the other events in the other three stages. We isolated a number of yeast mutants that weren't able to complete this cycle. We called them cell division cycle (cdc) mutants. Some cdc mutants turned out to have defects in the replication machinery. For example, cdc9 has a defective DNA ligase. DNA ligase is a protein that knits DNA pieces together. Because DNA polymerase adds nucleotides in the 5' to 3' direction, only one strand, the leading strand, is replicated as a continuous piece of DNA. The other strand, the lagging strand, is actually made in short 5' to 3' streches called Okazaki fragments. DNA ligase then knits the pieces together to make a continuous strand. cdc9 mutants have no functioning DNA ligase. The Okazaki fragments are never knitted together. Of course, it made sense that some cdc mutants had defects in the replication machinery. But we also found other types of cdc mutants. Normally, when exposed to radiation, cells stop in G1 to repair DNA damage before continuing onto the S stage. We isolated a mutant, rad9, that did not stop in G1 after radiation treatment. It continued on and finished the cell cycle by dividing. We believed that rad9 cells could no longer detect DNA damage caused by radiation. Thus, rad9 mutants divide and finish the cell cycle even when their DNA is not "ready." We tested this idea by making a double mutant that was both rad9 and cdc9. Remember, we knew that cdc9 had a defect in DNA ligase, so the DNA of one of the newly synthesized strands was in pieces. The double mutant finished the cell cycle by dividing even though the lagging strand was in pieces. Thus, rad9 works by stopping cells from dividing when DNA replication is incomplete or when DNA damage has occurred. We called these genes checkpoint genes. They allow cells to proceed to the next stage of the cell cycle only when specific requirements, like DNA synthesis and replication, have been met. We also isolated genes that acted as checkpoints for the other cell stages. Many of these checkpoint genes are also found in other species. Hi, I'm Bob Horvitz. I'm Mike Hengartner. Lee Hartwell told you about control mechanisms for cell growth and division. We’re going to tell you about control mechanisms for cell death. Mike and I work on an organism called Caenorhabitis elegans (C. elegans). It's a non-parasitic, microscopic roundworm with only 959 cells. The fate and lineage of every cell is known. This means that every cell division has been tracked so we know exactly where each of the 959 cells have come from and what it does. Not all organisms have a fixed cell lineage like this, which makes C. elegans development easier to study. The fertilized egg divides into two cells and so on to generate all the cells needed to make up the worm. Some of the cells in the lineages are programmed to die. The same cells always die at the same time in development. This phenomenon of programmed cell death or cell suicide is called apoptosis and is not the same as death due to injury or damage. Apoptosis is used as a way to clear superfluous, unwanted cells and is not unique to C. elegans. C. elegans actually generates 1090 cells during its development. It ends up with 959 cells because 131 are programmed to die. We can actually see cells dying. In this photo, the arrow points to a cell about to divide. One of its daughters will undergo programmed cell death. The cell from the previous photo has divided, and the arrows point to the two daughter cells. The division was unequal, and the smaller cell on the left will die. The dying cell refracts light differently and appears as a more distinct body. Neighboring cells will engulf the dead cell and get rid of the corpse. You can see the time lapse movie of this sequence of events in the AUDIO/VIDEO section of this concept. We became interested in the genes that carry out the cell death program. We called these genes cell death abnormals (ced). One in particular, ced-3, encodes a protein required for programmed cell death to occur. The ced-3 protein is a protease that actively degrades other proteins. In ced-3 mutants, cells that normally die don’t; they survive and often assume the function of their sister cells. Another gene, ced-9, encodes a protein that prevents programmed cell death in cells that should live. In other words, the ced-9 protein promotes cell survival. Mutations in ced-9 cause cells that normally live to die. The ced-3 and ced-9 proteins interact. Whenever it is present, the ced-3 protease will act on the ced-9 protein. This prevents ced-3 from killing the cells. In fact, the more ced-9 protein that is present, the better the protection against cell death. If there is enough ced-9 protein, then even cells that are supposed to die get protected against cell death. When we searched a gene database with the ced-3 and ced-9 sequences, we found two similar human genes: caspase-9 and bcl-2. These genes encode proteins with equivalent cell death functions in humans. In fact, we can replace the C. elegans ced-9 gene with human bcl-2 gene and still protect against cell death in C. elegans. Given the role of these genes in controlling cell growth and cell death, mutations in them can contribute to unchecked or unregulated growth leading to cancer. Hi, I'm Scott Lowe. I'm interested in the regulation of cell death in cancer. p53 is known to be a tumor-suppressor gene. Mice missing both copies of the p53 gene develop multiple malignant tumors. It turns out that, like the yeast rad9 gene, p53 is a checkpoint gene that monitors the state of the DNA. p53 protein levels rise after DNA damage, and the cells stop before the S stage. When DNA damage is extreme, high levels of p53 start the cell death program, and the cell undergoes programmed cell death. Again, bcl-2 acts as the protector and promotes cell survival. So, the cell death, repair and arrest pathways can be linked to tumor growth through the p53 tumor-suppressor gene.

Gallery


A young Leland Hartwell with his dog Sparky.
Leland Hartwell, President and Director of the Fred Hutchinson Cancer Research Center.
Robert Horvitz, Whitehead Professor of Biology, Massachusetts Institute of Technology.
Michael Hengartner, age three, with toy goat.
Michael Hengartner, age 18, in his high school chemistry lab.
Michael Hengartner with his kids, Alex and Sophie, 1999.
Michael Hengartner, Associate Professor, Watson School of Biological Sciences, Cold Spring Harbor Laboratory.
Scott Lowe working in his laboratory.

Audio/Video


Biography


 

Lee Hartwell was one of the first to use yeast as a model system, and he identified many of the genes involved in the cell cycle. Bob Horvitz and Mike Hengartner used C. elegans to work out the mechanism of programmed cell death. Scott Lowe's research has shown how the regulation of the cell cycle affect cancer.

LELAND HARTWELL (1939-)

Lee Hartwell was born in Los Angeles. His father was a sign maker, and Hartwell would help him in his shop after school. Hartwell developed an interest in figuring out how things work, especially electrical gadgets. This natural curiosity did not mean that he was a good student. He did well enough but didn't really get involved with science until after high school.

He was accepted to the California Institute of Technology and initially thought he would study physics. However, he became interested in "DNA" and finished college with a degree in biology. Hartwell then went on to earn a Ph.D. at the Massachusetts Institute of Technology.

After his Ph.D., Hartwell went to the Salk Institute because he wanted to work with Renato Dulbecco. Hartwell already knew he wanted to work on cell division, which was one of Dulbecco's research interests. Despite the cramped conditions - the Salk was a more primitive institute at the time - Hartwell enjoyed his post-doctoral years and learned a lot from his mentors.

Hartwell then accepted an assistant professorship at the University of California, Irvine. He also made the rather risky decision to start using yeast as a model system. Not many people were using yeast at the time, but Hartwell wanted and needed a simpler experimental system to study basic questions of cell growth. Hartwell is a pioneer in yeast genetics, and has used yeast to identify many of the genes involved in protein synthesis as well as the cell cycle.

In 1968, Hartwell moved to the Department of Genetics at the University of Washington and it was there that he did most of the work on cell cycle genes. He stayed at the University of Washington until 1996 when he moved to the Fred Hutchinson Cancer Research Center. His lab is working on the molecular mechanisms that maintain and support gene variations, which can eventually lead to the evolution of new species.

In 1997, Hartwell was appointed President and Director of the "Hutch," and spends most of his time integrating the basic, applied and clinical effort of interdisciplinary cancer research. In 1998, he won the Albert Lasker Basic Medical Research Prize for his innovative and pioneering work. Hartwell shared in the 2001 Nobel Prize in Physiology or Medicine for his work on defining the cell cycle.

HOWARD ROBERT HORVITZ (1947-)

Bob Horvitz was born in Chicago, Illinois. His mother was a teacher and his father was an accountant. Both parents instilled in Horvitz a respect and passion for learning. As a child, Horvitz maintained a butterfly collection, and thought that biology involved collecting and classifying dead things. He was a good student and interested in many subjects including English and journalism. He received two undergraduate degrees from the Massachusetts Institute of Technology, one in Mathematics and one in Economics. He considered going into law, medicine, business and even computer science. But because he didn't know anything about, and was intrigued by molecular biology (Horvitz didn't take any biology courses until his senior year in university) he entered graduate school at Harvard University to study biology.

Horvitz was interested in neurobiology, but because of his limited experience with biology in general, he started working with phage, to learn the basics. Horvitz was a graduate student in the laboratories of James Watson and Walter Gilbert, an experience he found "interesting." After his doctorate in 1974, Horvitz went to the Medical Research Council in Cambridge to work with Sydney Brenner.

Brenner was advocating a new model system for studying development. Caenorhabitis elegans is a non-parasitic roundworm that is amenable to genetic analysis, and is easy to grow and maintain. Horvitz saw the advantages of C. elegans, and used it to study a number of developmental systems including neuronal development, the ras pathway and the genetics of cell lineage. Programmed cell death is only one of the many ongoing projects in his lab.

In 1978, Horvitz accepted a position in the Department of Biology at the Massachusetts Institute of Technology. He is now Whitehead Professor of Biology. He has been an Howard Hughes Medical Institute investigator since 1988, and has won a number of awards for his work including the 1999 Gairdner Foundation Award. Horvitz has been a member of the National Academy of Science since 1991 and serves on a number of editorial boards and advisory committees, both governmental and commercial. He has a number of patents pending based on work done in his lab. Horvitz is a co-founder and Chairman of Idun Pharmaceuticals Inc., a biotech company based in La Jolla, California that is developing therapeutics focusing on apoptosis.

Horvitz enjoys reading in his spare time, particularly British contemporary novels.

Horvitz shared the 2002 Nobel Prize in Physiology or Medicine with colleagues John Sulston and Sdyney Brenner. All three made major contributions in the field of developmental biology using the model organism Caenorhabditis elegans.

MICHAEL OTMAR HENGARTNER (1966-)

Mike Hengartner was born in St. Gallin, Switzerland. His father was a professor of mathematics and moved his family from Switzerland to Paris, France, then Bloomington, Indiana before finally settling in Montreal, Canada. This globe-trotting at a young age gave Hengartner a facility for language. He speaks English, French and German.

Hengartner never had any doubts about being a scientist. His main problem was deciding which branch of science to focus on. Mathematics was out of the question because his older brother went into math. Hengartner initially thought of going into physics, but then read a book called What is Life? by Erwin Schrodinger. Hengartner realized that most physicists were switching fields and becoming biologists. Hengartner decided, therefore, not to waste time with physics. He graduated with a B.S. in biochemistry from Laval University in Quebec in 1988.

Hengartner was accepted into the Massachusetts Institute of Technology's biology department for graduate school. He was eager to work with Nobel Laureate David Baltimore on viral proteins. However, before making the final decision, Hengartner was persuaded by a friend to attend a lab meeting where he met Bob Horvitz who ran a Caenorhabiditis elegans lab. At the time, the C. elegans field was still rather new and Hengartner didn't really like the idea of working with worms. Yet, when Horvitz approached Hengartner to ask if he wanted to work in his lab, Hengartner was too "much of a coward" to say no. It did work out for the best because in a subsequent discussion about possible projects in the lab, Hengartner became fascinated with the idea of programmed cell death, and being able to determine the mechanism using C. elegans.

In 1994, Hengartner finished his doctorate in Horvitz's lab by cloning and characterizing ced-9, a gene necessary for programmed cell death in C. elegans.

Hengartner's work in Horvitz's lab led him to other genes involved in cell death both in C. elegans and in other organisms. After his Ph.D., Hengartner became a Staff Investigator at Cold Spring Harbor Laboratory (CSHL). He is currently an Associate Professor of the Watson School of Biological Sciences at CSHL. Hengartner also teaches at SUNY Stony Brook and organizes seminars and courses at CSHL and a number of other scientific institutions. He is an Executive Officer of the Cell Death Society and a cofounder of two biotech companies: Devgen, based in Belgium, and ForScience, based in New York. He is also on the editorial board of a number of science journals including Current Biology and Annals of Improbable Research. Hengartner has a number of patents pending based on his work.

Hengartner spends most of his leisure time with his family. He plays volleyball in the CSHL summer league, and believes that his poor game is due to his lack of height (Mike is 5'9"). Hengartner recently won the haiku portion of the 2000 Blackford Coffee Poetry Contest (Blackford is CSHL's cafeteria) with this submission:

Black oozy syrup

Brew'd daily by the gallons

Free but at what price?

SCOTT WILLIAM LOWE (1963-)

Scott Lowe was born in Racine, Wisconsin. Although he was good at science and took most of the courses in high school, he never was very interested in science. He actually thought he would become a lawyer.

In 1982, Lowe started at the University of Wisconsin-Madison in chemical engineering. He thought that chemical engineering had something to do with chemicals and engineering (math). He quickly found out that it wasn't the field for him, and decided to take some general courses to figure out what he did want to do. Lowe really became interested in biology and molecular genetics through his undergraduate biochemistry and genetics courses. As an undergraduate project, he went to work in a research lab, and after graduation stayed in the lab for two more years as a technician.

In 1988, Lowe went to the Department of Biology at the Massachusetts Institute of Technology to start his graduate work. Coincidentally, Michael Hengartner started graduate school on the same day in the same department. Little did Lowe know at the time, but his interest in oncogenes would one day overlap with Hengartner's interest in cell death genes.

After his Ph.D., Lowe stayed at MIT for post-doctorate work and began studying the effects a known tumor suppressor, p53, on cell growth. He found that in radiation-damaged cells, p53 is needed for programmed cell death. Connecting programmed cell death with tumor cell growth gave new insight as to how cancer cells proliferate.

In 1994, Lowe was offered a research position at Cold Spring Harbor Laboratory (CSHL). He is currently a professor at the Watson School of Biological Sciences at CSHL. Lowe's lab continues to work on the p53 pathway and the effects cell death genes have on the growth, proliferation and repair of tumor cells.

Lowe spends most of his free time with his family, especially his two young children. He claims to be able to sing the introduction to "Thomas the Tank Engine" backwards and forwards. He also likes to hike and camp and skiing is his favorite sport.

Factoid

Links


 

Links

Caenorhabditis elegans WWW server

A must for any worm researcher, this web site has resources and current information on worm research.

WormBase

A prototype, this web site will eventually contain all the sequences from the worm genome. It will be searchable and cross-referenced to mutations and known expression patterns.

Apoptosis

From the St. George's Hospital Medical School, this site has information on what happens within the cell during programmed cell death.

Bibliography

  • BioWeb, 1999, A Conversation/Lab Tour with H. Robert Horvitz, Harcourt College Publishers.

  • Clarke, A.R., et al., 1993, Thymocyte apoptosis induced by p53-dependent and independent pathways, Nature, 362: 849-852.

  • Elledge, S.J., 1996, Cell Cycle Checkpoints: Preventing an Identity Crisis, Science, 274: 1664-1672.

  • Hartwell, L.H. and Kastan, M.B., 1994, Cell Cycle Control and Cancer, Science, 266: 1821-1828.

  • Hartwell, L.H. and Weinert, T.A., 1989, Checkpoints: Controls That Ensure the Order of Cell Cycle Events, Science, 246: 629-634.

  • Hengartner, M.O. and Horvitz, H.R., 1994, C. elegans Cell Survival Gene ced-9 Encodes a Functional Homolog of the Mammalian Proto-Oncogene bcl-2, Cell, 76: 665-676.

  • Hengartner, M.O. and Horvitz, H.R., 1994, Programmed Cell Death in Caenorhabditis elegans, Current Opinion in Genetics and Development, 4: 581-586.

  • Lowe, S.W., Jacks, T., Housman, D.E., Ruley, H.E., 1994, Abrogation of oncogene-associated apoptosis allows transformation of p53-deficient cells, Proc. Natl. Acad. Sci., 91: 2026-2030.

  • Lowe, S.W., Schmitt, E.M., Smith, S.W., Osborne, B.A. and Jacks, T., 1993, p53 is required for radiation-induced apoptosis in mouse thymocytes, Nature, 362: 847-849.

  • Murray, A., 1994, Cell Cycle Checkpoints, Current Opinion in Cell Biology, 6: 872-876.

  • Spector, M.S., Desnoyers, S., Hoeppner, D.J., Hengartner, M.O., 1997, Interaction between the C. elegans cell-death regulators CED-9 and CED-4, Nature, 385: 653-656.

  • Xue, D. and Horvitz, H.R., 1997, Caenorhabditis elegans CED-9 protein is a bifunctional cell-death inhibitor, Nature, 390: 305-308.

Glossary



Children resemble their parents.
Genes come in pairs.
Genes don't blend.
Some genes are dominant.
Genetic inheritance follows rules.
Genes are real things.
All cells arise from pre-existing cells.
Sex cells have one set of chromosomes; body cells have two.
Specialized chromosomes determine gender.
Chromosomes carry genes.
Genes get shuffled when chromosomes exchange pieces.
Evolution begins with the inheritance of gene variation.
Mendelian laws apply to human beings.
Mendelian genetics cannot fully explain human health and behavior.
DNA and proteins are the molecules of the cell nucleus.
One gene makes one protein.
A gene is made of DNA.
Bacteria and viruses have DNA too.
The DNA molecule is shaped like a twisted ladder.
A half DNA ladder is a template for copying the whole.
RNA is an intermediary between DNA and protein.
DNA words are three letters long.
A gene is a discrete sequence of DNA nucleotides.
The RNA message is sometimes edited.
Some viruses store genetic information in RNA.
RNA was the first genetic molecule.
Mutations are changes in genetic information.
Some types of mutations are automatically repaired.
A chromosome is a package for DNA.
Higher cells incorporate an ancient chromosome.
Some DNA does not encode protein.
Some DNA can jump.
Genes can be turned on and off.
Genes can be moved between species.
DNA responds to signals from outside the cell.
Different genes are active in different kinds of cells.
Master genes control basic body plans.
A genome is an entire set of genes.
Living things share common genes.
DNA is only the starting point for understanding human biology.
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