Slides 04 Viruses MicrobiologyAdvancedddb

Viruses
With Prof. Vincent Racaniello
Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

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Learning Goals
• You will know how viruses are cultivated and assayed.
• You will distinguish the seven different types of viral

genomes and their structures.
• You will appreciate what information is and is not

encoded in viral genomes.

How Are Viruses Studied?
• At first, animal viruses could not be routinely

propagated in cell cultures
• Most viruses were grown in laboratory animals

Studying the Infectious Cycle in Cells
1950 1955
Studying not possible before
1949 (animal viruses)

1950 1955
1949 Enders, Weller, Robbins propagate

poliovirus in human cell culture – primary
cultures of embryonic tissues

1950 1955
1954 Nobel Prize

Today, Viruses Are Routinely Propagated in Cell Cultures
Primary human Mouse fibroblast Human epithelial

foreskin fibroblasts cell line (3T3) cell line (HeLa)
Continuous cell lines
Diploid cell strains (e.g. WI-38, human embryonic lung)

Growing Cells in Culture

Detecting Viral Multiplication – Cytopathic Effects

Detecting Viruses in Samples
Infectivity
Physical: virus particles

and their components

Detecting Viruses in Samples
Infectivity
Physical: virus particles

and their components

Detecting Viruses by Plaque Assay
0.1 ml 0.1 ml
0.9 ml
Virus
stock
10–1 10–2 10–3 10–4 10–5 10–6 10–7
Number of Plaques: Too many to count 17 2

1.7 x 108 PFU/ml
The Wall of Polio

Physical Measurements of Virus Particles
Hemagglutination Electron microscopy Viral enzymes
Serology Nucleic acids

Next Generation DNA Sequencing
• Deep, high-throughput
sequencing
• Identification of new viruses

in environmental samples
• Identification of new
pathogens

Frigid Antarctica is loaded with viruses
• Deep, high-throughput
sequencing Researchers found
nearly 10,000
• Identification of new viruses species by
in environmental samples analyzing water
samples
pathogens

TWiV 196: An arena for snakes

AUGUST 19, 2012
Hosts: Vincent Racaniello, Alan

Dove, Rich Condit, Dickson
• Deep, high-throughput Despommier, Kathy Spindler, Mark

Stenglein, and Josep DeRisi
sequencing The TWiVites meet with Mark
Stenglein and Joseph de Risi
• Identification of new viruses to discuss their discovery of a
in environmental samples novel arenavirus in snakes
with inclusion body disease.
pathogens

Virology Breakthrough in the 1950s
The viral nucleic acid genome is the genetic code

Virology Breakthrough in the 1950s
Hershey-Chase experiment with phage T4 Fraenkel-Conrat’s work with TMV

The Bigger Surprise
Despite the countless numbers of viruses on Earth,

there is a finite number of viral genomes

A Key Fact That Makes Life Easier for Students of Virology
• Viral genomes must make mRNA that

can be read by host ribosomes
• All viruses on the planet follow this rule.

There is no known exception.

Nobel Laureate David Baltimore Used This Insight to Describe a Simple
Way to Think About Virus Genomes
+ DNA II
VI ± DNA
VII
+ RNA – DNA ± DNA I
+ RNA – RNA + mRNA ± RNA

IV III
– RNA V
The Baltimore system
Knowing only the nature of the viral genome, one can deduce
the basic steps that must take place to produce mRNA

Definitions
+ DNA II
• mRNA (ribosome ready) is
always the (+) strand VI ± DNA
VII
• DNA of equivalent polarity is + RNA – DNA ± DNA I
also the (+) strand
• RNA and DNA complements of (+) + RNA – RNA + mRNA ± RNA

strands are negative (-) strands IV III
• Not all (+) RNA is mRNA!
– RNA V

What Information is Encoded in a Viral Genome?
Gene products and regulatory signals for: Ori Ori

ITR
3‘ 5‘
Replication of the viral genome 5‘ 3‘
ITR
Assembly and packaging of the genome
Regulation and timing of the replicative cycle
Modulation of host defenses ± DNA ± DNA
Spread to other cells and hosts


ITR
3‘ 5‘
ITR
Modulation of host defenses


ITR
3‘ 5‘
ITR


ITR
3‘ 5‘
ITR


ITR
3‘ 5‘
ITR

Information NOT Encoded in Viral Genomes
No genes encoding the complete protein

NO synthesis machinery (AARS, eIFs, tRNAs)
No genes encoding proteins involved in

NO energy production or membrane
biosynthesis
NO No classical centromeres or telomeres

found in standard host chromosomes
100 nm

Information NOT Encoded in Viral Genomes
We probably haven‘t
found them yet –
most giant virus
genes are
novel ?
?
? ?
?
Largest Known Viral Genomes
Virus Length Protein

Pandoravirus salinus 2,473,870 2,541
Pandoravirus dulcis 1,908,524 1,487
Megavirus chilensis 1,259,197 1,120
Mamavirus 1,191,693 1,023
Mimivirus 1,181,549 979
Moumouvirus 1,021,348 894
Mimivirus M4 981,813 620
C. roenbergensis virus 617,453 544
Mollivirus sibericum 651,000 523
Pithovirus sibericum 610,033 467

Smallest Known Viral Genomes
Virus Length Protein

Viroid 120 none
Satellite 220 none
Hepatitis delta satellite 1,700 1
Circovirus 1,759 2
Anellovirus 2,170 4
Geminivirus 2,500 4
Hepatitis B virus 3,200 7
Levivirus 3,400 4
Partitivirus 3,700 2
Barnavirus 4,000 7

Learning Outcomes
 You will know how viruses are cultivated and assayed.
 You will understand the seven different types of viral

genomes and their structures.
 You will appreciate what information is and is not

encoded in viral genomes.

Dieses Dokument gehört: Marcos Santiago Campos Rosas
Hinweis: Dieses Dokument ist urheberrechtlich geschützt. Die Vervielfältigung,

Verbreitung, Wieder- bzw. Weitergabe und sonstige Nutzung ist ohne die schriftliche
Genehmigung der Lecturio GmbH nicht gestattet.


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Viruses

With Prof. Vincent Racaniello

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

• You will know how viruses are cultivated and assayed.

• You will distinguish the seven different types of viral

• You will appreciate what information is and is not

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

• At first, animal viruses could not be routinely

• Most viruses were grown in laboratory animals

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Studying not possible before

1949 (animal viruses)

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

1949 Enders, Weller, Robbins propagate

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

1954 Nobel Prize

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Primary human Mouse fibroblast Human epithelial

Continuous cell lines

Diploid cell strains (e.g. WI-38, human embryonic lung)

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Physical: virus particles

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Physical: virus particles

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Number of Plaques: Too many to count 17 2

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Hemagglutination Electron microscopy Viral enzymes

Serology Nucleic acids

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

• Identification of new viruses

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Frigid Antarctica is loaded with viruses

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

TWiV 196: An arena for snakes

Hosts: Vincent Racaniello, Alan

• Deep, high-throughput Despommier, Kathy Spindler, Mark

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

The viral nucleic acid genome is the genetic code

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Hershey-Chase experiment with phage T4 Fraenkel-Conrat’s work with TMV

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Despite the countless numbers of viruses on Earth,

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

• Viral genomes must make mRNA that

• All viruses on the planet follow this rule.

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

+ RNA – RNA + mRNA ± RNA

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

• RNA and DNA complements of (+) + RNA – RNA + mRNA ± RNA

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Gene products and regulatory signals for: Ori Ori

Regulation and timing of the replicative cycle

Modulation of host defenses ± DNA ± DNA

Spread to other cells and hosts

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com

Gene products and regulatory signals for: Ori Ori

Regulation and timing of the replicative cycle

Modulation of host defenses

Spread to other cells and hosts

Marcos Santiago Campos Rosas, marcosantiago896@gmail.com