Chapter 18

1. Chapter 18 The Genetics of Virusesand Bacteria

2. Concept 18.1: A virus has a genome but can reproduce only within a host cell • Scientists detected viruses indirectly long before they could see them • The story of how viruses were discovered begins in the late 1800s

3. The Discovery of Viruses: Scientific InquiryTobacco mosaic disease • Tobacco mosaic disease stunts the growth of tobacco plants and gives their leaves a mottled, or mosaic, coloration .

5. In 1883, Adolf Mayer, a German scientist, discovered that he could transmit the disease from plant to plant by rubbing sap extracted from diseased leaves onto healthy plants. • After an unsuccessful search for an infectious microbe in the sap, Mayer concluded that the disease was caused by unusually small bacteria that could not be seen with the microscope.

7. This hypothesis was tested a decade later by Dimitri Ivanowsky, a Russian who passed sap from infected tobacco leaves through a filter designed to remove bacteria. • After filtering, the sap still produced mosaic disease • Ivanowsky clung to the hypothesis that bacteria caused tobacco mosaic disease. • Perhaps, he reasoned, the bacteria were so smallthat they passed through the filter or made a filterable toxin that caused the disease

8. Dutch botanist Martinus Beijerinck discovered that the infectious agent in the filtered sap could reproduce.

9. In fact, the pathogen reproduced only within the host it infected… • Unlike bacteria, the mysterious agent of mosaic disease couldnot be cultivated on nutrient media in test tubes or petri dishes. • Beijerinck imagined a reproducing particle much smaller and simpler than bacteria. • His suspicions were confirmed in 1935 when the American scientist Wendell Stanley crystallized the infectious particle, now known as tobacco mosaic virus (TMV). Subsequently, TMV and many other viruses were actually seen with the help of the electron microscope.

10. The Discovery of Viruses: Scientific Inquiry • Tobacco mosaic disease stunts growth of tobacco plants and gives their leaves a mosaic coloration • In the late 1800s, researchers hypothesized that a particle smaller than bacteria caused the disease • In 1935, Wendell Stanley confirmed this hypothesis by crystallizing the infectious particle, now known as tobacco mosaic virus (TMV)

11. Structure of Viruses • The tiniest viruses are only 20 nm in diameter— smaller than a ribosome. • Stanley′s discovery that some viruses could be crystallized was exciting and puzzling news. • Not even the simplest of cells can aggregate into regular crystals. • But if viruses are not cells, then what are they?

12. They are infectious particles consisting of nucleic acid enclosed in a protein coat and, in some cases, a membranous envelope.

13. Viral Genomes • We usually think of genes as being made of double–stranded DNA—the conventional double helix—but many viruses defy this convention. • Their genomes may consist of double–stranded DNA, single–stranded DNA, double–stranded RNA, or single–stranded RNA, depending on the kind of virus. • A virus is called a DNA virus or an RNA virus, according to the kind of nucleic acid that makes up its genome. In either case, the genome is usually organized as a single linear or circular molecule of nucleic acid. The smallest viruses have only four genes, while the largest have several hundred

14. Capsids and Envelopes • The protein shell enclosing the viral genome is called a capsid. • Depending on the type of virus, the capsid may be rod–shaped, polyhedral, or more complex in shape (like T4). • Capsids are built from a large number of protein subunits called capsomeres, but the number of different kinds of proteins is usually small.

15. Tobacco mosaic virus has a rigid, rod–shaped capsid made from over a thousand molecules of a single type of protein arranged in a helix .

16. Adenoviruses, which infect the respiratory tracts of animals, have 252 identical protein molecules arranged in a polyhedral capsid with 20 triangular facets—an icosahedron .

17. Some viruses have accessory structures that help them infect their hosts. For instance, a membranous envelope surrounds the capsids of influenza viruses and many other viruses found in animals.

18. These viral envelopes, which are derived from the membrane of the host cell, contain host cell phospholipids and membrane proteins. • They also contain proteins and glycoproteins of viral origin (glycoproteins are proteins with carbohydrate covalently attached). • Some viruses carry a few viral enzyme molecules within their capsids.

19. Bacteriophages • The most complex capsids are found among viruses that infect bacteria, called bacteriophages, or simply phages.

20. The first phages studied included seven that infect E. coli. • These seven phages were named type 1 (T1), type 2 (T2), and so forth, in the order of their discovery. • The three T–even phages (T2, T4, and T6) turned out to be very similar in structure. • Their capsids have elongated icosahedral heads enclosing their DNA. • Attached to the head is a protein tail piece with fibers that the phages use to attach to a bacterium.

21. General Features of Viral Reproductive Cycles • Viruses are obligate intracellular parasites: • They can reproduce only within a host cell. An isolated virus is unable to reproduce or do anything else except infect an appropriate host cell. • Viruses lack metabolic enzymes, ribosomes, and other equipment for making proteins. • Thus, isolated viruses are merely packaged sets of genes in transit from one host cell to another

22. Host specificity • Each type of virus can infect only a limited range of host cells, called its host range. • This host specificity results from the evolution of recognition systems by the virus. • Viruses identify their host cells by a “lock–and–key” fit between proteins on the outside of the virus and specific receptor molecules on the surface of cells. • Some viruses have broad host ranges.

23. West Nile virus, for example, can infect mosquitoes, birds, and humans, and equine encephalitis virus can infect mosquitoes, birds, horses, and humans. • Other viruses have host ranges so narrow that they infect only a single species. • Measles virus and poliovirus, for instance, can infect only humans. • Furthermore, infection by viruses of multicellular eukaryotes is usually limited to particular tissues. • Human cold viruses infect only the cells lining the upper respiratory tract, and the AIDS virus binds to specific receptors on certain types of white blood cells.

24. A viral infection • A viral infection begins when the genome of a virus makes its way into a host cell. • How? • What is the mechanism?

25. The mechanism by which this nucleic acid enters the cell varies, depending on the type of virus and the type of host cell. • For example, the T–even phages use their elaborate tail apparatus to inject DNA into a bacterium. • Once inside, the viral genome can commandeer its host, reprogramming the cell to copy the viral nucleic acid and manufacture viral proteins.

26. The host provides… • the nucleotides for making viral nucleic acids, as well as enzymes, ribosomes, tRNAs, amino acids, ATP, and other components needed for making the viral proteins dictated by viral genes.

27. DNA viruses vs RNA viruses • Most DNA viruses use the DNA polymerases ofthe host cellto synthesize new genomes along the templates provided by the viral DNA. • In contrast, to replicate their genomes, RNA viruses use special virus–encoded polymerases that can use RNA as a template. • (Uninfected cells generally make no enzymes for carrying out this latter process.)

28. Overview: Microbial Model Systems • Viruses called bacteriophages can infect and set in motion a genetic takeover of bacteria, such as Escherichia coli • E. coli and its viruses are called model systems because of their frequent use by researchers in studies that reveal broad biological principles • Beyond their value as model systems, viruses and bacteria have unique genetic mechanisms that are interesting in their own right

29. A simplified viral reproductive cycle. • A virus is an obligate intracellular parasite that uses the equipment and small precursors of its host cell to reproduce. In this simplest of viral cycles, the parasite is a DNA virus with a capsid consisting of a single type of protein

30. LE 18-5 VIRUS Entry into cell and uncoating of DNA DNA Capsid Transcription Replication HOST CELL Viral DNA mRNA Viral DNA Capsid proteins Self-assembly of new virus particles and their exit from cell

31. assembly of viruses… • After the viral nucleic acid molecules and capsomeres are produced, their assembly into new viruses is often a spontaneous process of self–assembly. • In fact, the RNA and capsomeres of TMV can be separated in the laboratory and then reassembled to form complete viruses simply by mixing the components together under the right conditions. • The simplest type of viral reproductive cycle ends with the exit of hundreds or thousands of viruses from the infected host cell, a process that often damages or destroys the cell.

32. Such cellular damage and death, as well as the body′s responses to this destruction, cause some of the symptoms associated with viral infections. • The viral progeny that exit a cell have the potential to infect additional cells, spreading the viral infection. • There are many variations on the simplified viral reproductive cycle. • We will now take a closer look at some of these variations in bacterial viruses (phages) and animal viruses;later in the chapter, we will consider plant viruses

33. Reproductive Cycles of Phages • Phages are the best understood of all viruses, although some of them are also among the most complex. • Research on phages led to the discovery that some double–stranded DNA viruses can reproduce by two alternative mechanisms: • the lytic cycle and • the lysogenic cycle.

34. The Lytic Cycle • A phage reproductive cycle that culminates in death of the host cell is known as a lytic cycle. • The term refers to the last stage of infection, during which the bacterium lyses (breaks open) and releases the phages that were produced within the cell. • Each of these phages can then infect a healthy cell, and a few successive lytic cycles can destroy an entire bacterial population in just a few hours. • A phage that reproduces only by a lytic cycle is a virulent phage

35. Figure 18.6   The lytic cycle of phage T4, a virulent phage.  • Phage T4 has about 100 genes, which are transcribed and translated using the host cell′s machinery.

36. One of the first phage genes translated after the viral DNA enters the host cell codes for an enzyme that degrades the host cell′s DNA (step 2

37. the phage DNA is protected from breakdown because it contains a modified form of cytosine that is not recognized by the enzyme.

38. The entire lytic cycle, from the phage′s first contact with the cell surface to cell lysis, takes only 20–30 minutes at 37°C.

39. LE 18-6 Attachment Entry of phage DNA and degradation of host DNA Phage assembly Release Head Tail fibers Tails Synthesis of viral genomes and proteins Assembly

41. Bacteria are prokaryotes with cells much smaller and more simply organized than those of eukaryotes • Viruses are smaller and simpler than bacteria

42. LE 18-2 Virus Bacterium Animal cell Animal cell nucleus 0.25 µm

43. why phages haven′t exterminated all bacteria? • Bacteria are not defenseless. • First, natural selection favors bacterial mutants with receptor sites that are no longer recognized by a particular type of phage. • Second, when phage DNA successfully enters a bacterium, the DNA often is recognized as foreign and cut up by cellular enzymes called restriction endonucleases, or simply restriction enzymes. The bacterial cell′s own DNA is chemically modified in a way that prevents attack by restriction enzymes. • There is yet a third important reason is that instead of lysing their host cells, many phages coexist with them in what is called the lysogenic cycle

44. In fact, phage treatments have been used medically in some countries to help control bacterial infections.

45. evolutionary flux… • But just as natural selection favors bacteria with effective restriction enzymes, natural selection favors phage mutants that are resistant to the bacterial enzymes. • Thus, the parasite–host relationship is in constant evolutionary flux.

46. The Lysogenic Cycle • In contrast to the lytic cycle, which kills the host cell, the lysogenic cycle replicates the phage genome without destroying the host. • Phages capable of using both modes of reproducing within a bacterium are called temperate phages. • A temperate phage called lambda, written with the Greek letter λ, is widely used in biological research. • Phage λ resembles T4, but its tail has only one short tail fiber.

47. LE 18-7 Phage DNA The phage attaches to a host cell and injects its DNA. Daughter cell with prophage Many cell divisions produce a large population of bacteria infected with the prophage. Phage DNA circularizes Phage Bacterial chromosome Occasionally, a prophage exits the bacterial chromosome, initiating a lytic cycle. Lytic cycle Lysogenic cycle The bacterium reproduces normally, copying the prophage and transmitting it to daughter cells. Certain factors determine whether The cell lyses, releasing phages. Lytic cycle is induced Lysogenic cycle is entered or Prophage Phage DNA integrates into the bacterial chromosomes, becoming a prophage. New phage DNA and proteins are synthesized and assembled into phages.

48. Infection of an E. coli cell by phage λ • Infection begins when the phage binds to the surface of the cell and injects its DNA. • Within the host, the λ DNA molecule forms a circle. • What happens next depends on the reproductive mode: lytic cycle or lysogenic cycle.

49. lytic cycle • During a lytic cycle, the viral genes immediately turn the host cell into a λ–producing factory, and the cell soon lyses and releases its viral products.

50. lysogenic cycle • During a lysogenic cycle, however, the λ DNA molecule is incorporated by genetic recombination (crossing over) into a specific site on the host cell′s chromosome.