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Virus Diversity
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Micr221: Virology Section

Instructor:

Dr. Eric Carstens

there is a Moodle site read the FAQs (frequently asked questions) from 2003 and later post all questions to Questions About Virology link on the Online Course Moodle page.

if you send me questions about course material by email, other students dont get to see the responses!

ofce hours Mondays 16:00 -17:00 hr room 714 Botterell Hall

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Textbook: Virology Sections

Recommended: Microbiology 8th (7th or 6th similar) (Prescott, Harley, Klein) (on reserve in Bracken Library)

Review Part Four chapter 12:

Genes: structure, replication, expression Recombinant DNA technology/Genomics Part One Chapter 5

9th Edition is not as good for virology

Review Part Four chapter 15 and 16 Learn Part One and Five The Viruses

general properties, structure, culturing, studying taxonomy and replication strategies

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Part Five Chapter 25

Learning Expectations

Part Eight, Chapter 37 supplemental to lecture material:

human diseases caused by viruses and prions

as mentioned in lectures (I am going to focus on the viruses and not their effects on humans or as causative agents of disease).

Learn:

material about viruses discussed in lectures Classication of Viruses 7th ed (on Moodle under the heading Taxonomy of Viruses) ViralZone

Excellent web sites:

expasy.org/viralzone www.ictvonline.org/virusTaxonomy.asp?version=2011

Virus Database On-line (ICTVdb):

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Excellent Online Resource

Fields Virology [electronic resource] / editors-in-chief, David M. Knipe, Peter M. Howley ; associate editors, Diane E. Grifn ... [et al.].

Queen's access only http://gateway.ovid.com/ovidweb.cgi? T=JS&NEWS=N&PAGE=booktext&D=books&AN=00 139921/5th_Edition/5&XPATH=/OVIDBOOK %5b1%5d/METADATA%5b1%5d/TBY%5b1%5d/ EDITORS%5b1%5d

Overview of Virology Lecture Series

Topical slide to start each lecture History

Discovery and isolation of viruses What is a virus?

Virus structure and taxonomy Principles of virus replication - steps in the replication cycle Classes of viruses and their modes of replication Viruses as tools in molecular virology - expression vectors, virus-host interactions
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Lecture Overheads

HOPEFULLY, WHAT I SAY IS IMPORTANT TO YOUR LEARNING! Slide Title will usually provide essential information You are responsible for:

material discussed in class material in text book related to what is discussed in class knowing characteristics of major families of viruses discussed during the lectures

lecture slides are available on Moodle as PDFs (if you want a different format, let me know)
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By the end of this section, you should be able to:

identify major virus families and understand their structure and replication strategies know how information about viruses is derived recognize the role viruses play in life sciences (knowledge generation) in addition to their potential as pathogens go out into the world and critically analyze articles and reports about viruses

Background to the Discovery of Viruses

Louis Pasteur: 1822-1895 Robert Koch: 1843-1910 Joseph Lister: 1827-1912

Their discoveries lead to a new experimental approach for medical science, dening whether an organism was the causative agent of a disease
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Kochs Postulates

1. The same pathogen must be present in every case of the disease 2. The pathogen must be isolated from the diseased host and grown in pure culture 3. The pathogen from the pure culture must cause the disease when inoculated into a healthy, susceptible laboratory animal 4. The pathogen must be isolated from the inoculated animal and must be shown to be the original organism The Germ Theory as proposed by Pasteur and articulated by Koch (and his postulates) did not recognize the existence of viruses.

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What is a virus?

In the beginning ...

Agent of Disease or Symptoms

Adolf Mayer (1843-1942), German working in Holland tobacco mosaic disease light and dark spots on leaves juice from infected leaves inoculated into healthy plants caused disease transmission agent was unknown, speculated it was caused by an unknown infectious form
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Filterable Infectious Agent

Dimitri Ivanofsky (1864-1920) commissioned by Russian Department of Agriculture tobacco disease repeated Mayers experiments, with additional step

passed infected sap through a lter known to block bacteria 1892 - the sap of leaves infected with tobacco mosaic disease retains its infectious properties even after ltration through Chamberland lter candles

provided operational denition of viruses - The concept of lterable infectious agent was against all accepted scientic concepts at
that time, so it was not widely accepted and Iwanowski himself even suspected the disease was a result of a toxin.
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Chamberland Filter
The Berlin Inquiry of 1886 gave great prominence to the filtration of water for domestic purposes, and that made for the German War Office in 1895 by Dr. Plagge drew particular attention to the Pasteur Chamberland The Pasteur Chamberland filter is made of a porous porcelain tube through which the water is forced under pressure. The residue left on the outside of the tubes can easily be removed, and the tubes themselves should be sterilized periodically by boiling. At Darjeeling 9500 of these tubes are in use in the municipal water-works, and the supply given is 150,000 gallons a day.
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Obligate Intracellular Parasite

Martinus Beijerinck (1851-1931), collaborated with Mayer ltered sap could be diluted and then regain its strength (virulence) after transfer through living, growing plant tissue

contagium vivum uidum (soluble living germ)

agent remained infective through several transfers so was not a toxin explained failure to culture the pathogen outside its host

1998 centenary celebration of virology (Tobacco mosaic virus) (TMV) http://www.sciencedirect.com/science/article/pii/ S0966842X98013754
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Original colour illustrations for Beijerinck's paper about TMV ~1898

http://www.tnw.tudelft.nl/live/pagina.jsp?id=8e07d591c3c7-4801-94c2-82302a62be90&lang=en

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Obligate Intracellular Parasite (cont.)

new Concept for the turn of the 20th century:

a lterable agent too small to observe in the light microscope but able to cause disease by multiplying in living cells

Loefer and Frosch (1898) isolated rst lterable agent from animals (Foot-and-mouth disease virus - family Picornaviridae) Reed et al. (1900) recognized rst human lterable agent (Yellow fever virus - family Flaviviridae) and also showed mosquito transmission of agent

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Chemical makeup of lterable agents

methods to purify proteins (precipitation) also puried Tobacco mosaic virus (TMV)

how did they detect the agent?

assayed by infectivity (in host plants!)

TMV, like proteins, migrated in an electric eld rabbit antibodies directed against TMV neutralized infectivity Led to the question:

Are viruses proteins?

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Viruses contain both protein and nucleic acid

by 1929, concluded that viruses consisted of protein but, in 1934, Max Schlesinger in Germany showed that bacteriophages contained both protein and deoxyribonucleic acid (DNA)

rst suggestion that viruses were composed of nucleoprotein (nucleic acid plus protein)

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Viruses contain only one kind of nucleic acid

in 1935, Wendell Stanley crystallized TMV

these infectious crystals contained protein and 5% RNA

in 1940, Hoagland at the Rockefeller Institute showed that vaccinia virus contained DNA but no RNA Concept:

Virus genetic material is DNA or RNA but not both!

TMV crystals
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Viruses contain only one kind of nucleic acid

in 1935, Wendell Stanley crystallized TMV

these infectious crystals contained protein and 5% RNA

in 1940, Hoagland at the Rockefeller Institute showed that vaccinia virus contained DNA but no RNA Concept:

Virus genetic material is DNA or RNA but not both!

TMV crystals
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What do viruses look like?

Helmut Ruska (c 1969)

in 1939, the rst electron micrographs showed TMV was rod shaped while x-ray diffraction suggested TMV was built up from repeating subunits. Concept:

most simple viruses consist of one or a few identical protein subunits

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What do viruses look like?

Helmut Ruska (c 1969)

in 1939, the rst electron micrographs showed TMV was rod shaped while x-ray diffraction suggested TMV was built up from repeating subunits. Concept:

most simple viruses consist of one or a few identical protein subunits

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Electron Micrographs of Some Different Viruses

Adenovirus

Rhabdovirus

Herpesvirus

Tobacco mosaic virus Coronavirus Papillomavirus

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So ... what is a virus?

a virus is an infectious, obligate intracellular parasite the genetic material of a virus enters a host cell and directs the production of the building blocks of new virus particles (called virions) new virions are made in the host cell by assembly of these building blocks the new virions produced in a host cell then transport the viral genetic material to another host cell or organism to carry out another round of infection

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So ... what is a virus?

viruses differ from bacteria in size, their inability to grow in lifeless media and that they contain only one kind of nucleic acid, either DNA or RNA lterable, disease-causing agent, completely dependent upon living cell (intracellular parasite)

self replicating in host cell inert (dormant) outside a living cell

contrasts with dynamic growth of bacteria

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Why memorize virus facts?

viruses are easy to understand when we reduce their properties to simple descriptions such as those listed above. The confounding issues lie in the details, and with viruses, there are many, many details. knowledge comes from understanding the details

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Viruses are Everywhere!

Inside a typical ofce building there are 400-900 microorganisms per cubic meter of air. The majority of these are viruses. With a resting ventilation rate of ~6 liters/minute, the average adult would inhale about 10,000 micro-organisms per day! Why arent we always sick?

Host defenses: physical and immunological (innate and adaptive immunity). Microbial specicity: proportionally, very few microbes are capable of infecting us. Inapparent infections: often we are infected but with no overt signs.
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Did you know? Viruses and the Sea

Best estimates range from ~3 x 106 viruses/ml in the deep sea to ~108/ml in productive coastal waters. The oceans contain a total of ~4 x 1030 viruses equal to 200 Megatonnes (200 x 109 kg) of carbon, roughly the amount of carbon in 75 million blue whales! If aligned end to end they would span ~10 million light years, or ~100 times the distance across our galaxy! Every second ~1023 viral infections occur in the ocean. Viruses are the most abundant biological entity in the oceans and the second largest component of biomass after prokaryotes.
Sources: C. A. Suttle. Nature Reviews Microbiology. 2007. 5:801. C. A.Suttle (2005).Viruses in the sea. Nature 437:356.

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Why Study the Molecular Biology of Viruses?

There is more biological diversity within viruses than in all the rest of bacterial, plant and animal kingdoms put together! Virus/host interactions are molecular negotiations of fundamental and practical signicance that must be understood if we are to defend ourselves against viruses. However, the very things that make viruses insidious and lethal can be exploited for practical use through the development of viruses as productive tools and therapeutic agents.

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Questions?