Viruses Defined by Unique Properties

ÎObligate intracellular parasites that do not

undergo binary fission
6 distinguish from bacterial pathogens and other
parasites

ÎParasitic genomes related to plasmids that can

exist in two forms
6 distinguish from other life forms
 Virus Life Cycle
Extracellular (metabolically inert) Intracellular (replicating)
Entry/Penetration*
Attachment*

genome
uncoated
Synthesis
mRNA replication &
(RNA = cytoplasm Replication
DNA = nucleus)
proteins

progeny virion Assembly

Release*
Virus particle (virion)
1. Genome (DNA or RNA)
2. Protective coat (capsid)
with or without envelope
3. Transport, attachment,
facilitate penetration
 Definitions
Virus Particle = VIRION
• Naked: coat (capsid) protein (CP) + genome

• Enveloped: Lipid bilayer membrane surrounds encapsidated

genome (nucleocapsid)
Envelope acquired from host cell membranes (at site
of virus assembly)

Genome = DNA or RNA (never both!)

• Single-stranded (ss) or double-stranded (ds)

• ssRNA viruses classified by the polarity of their genome

ƒ Positive-sense (+) = polarity of mRNA
ƒ Negative-sense (–) = opposite polarity to mRNA
 Plant Virus Life Cycle: Adaptations to the Cell Wall at
Entry and Exit
entry/penetration*
Attachment/Penetration:
animal viruses bind to specific
surface receptors;
Entry: fuse with or engulfed by
genome
the plasma membrane
uncoated Plant viruses: mechanical
introduction initially. NO evidence
(RNA = cytoplasm attach to specific receptor sites on
mRNA replication DNA = nucleus) cell wall

Transmission:
animal viruses: aerosols, break in
proteins skin, fluids [blood, sexual contact]
plant viruses: insects, fungi,
nematodes, abrasion, seeds,
pollen, vegetative propagation
progeny virion assembly
Release:
animal viruses lyse cells or bud
through (plasma) membrane
plant viruses “channel” through
“release” * wall (MPs) without lysis
 There is no Evidence that Plant Viruses Attach to
Specific Receptor Sites on Plant Cells

The stages of plant virus penetration and uncoating of the viral

genome are not understood

Plant viruses are initially introduced through the cell wall by

mechanical means when a plant is first infected

Studies with some plant viruses (e.g. TMV, TBSV) suggest that bound
Ca+2 can stabilize virion capsid structure

It has been suggested that the low Ca+2 concentrations inside plant
cells may facilitate uncoating of at least some plant viruses
 TABLE 2. Transmission of Viruses Between Hosts1

AEROSOLS FLUIDS PARENT TO VECTORS

(airborne) OR (direct OFFSPRING
INGESTION contact)
(water- or
foodborne)
MOST FEW FEW MANY
ANIMAL Picorna Hepadna Retro Toga
VIRUSES Orthomyxo Retro Herpes Flavi
Corona Herpes Arena Bunya
Reo Papilloma Rhabdo

NONE FEW MANY MOST

PLANT Tobamo Hordei Poty
VIRUSES Tombus Ilar Potex
[Water, Poty Gemini
Mechanical] [Seeds, Luteo
Pollen, Bulbs, Tospo (Bunya)
Cuttings] [Insects,
Nematodes, Fungi]
1
Examples indicate that some, not necessarily all, viruses within the family are transmitted by the
given mode.

Lazarowitz (2001) in Fundamental Virology (Knipe & Howley, eds. )4th Ed, Lippincott
 Virus Intercellular Transport
The goal is for progeny viruses to infect new host cells and repeat
the infectious cycle
Í May be an immediate neighbors or a distant cells
Í In animals and humans, the circulatory and nervous systems are
conduits for systemic infection (dissemination to distant sites)

Most Animal Viruses go through an extracellular form (released virions)

to infect additional cells, even locally
Í Local infection will occur through interstitial or extracellular
fluids (e.g. respiratory viruses will spread through mucus)

All Plant Viruses spread directly cell to cell within a leaf without an
extracellular phase (local movement)
Í An extracellular form is transported leaf to leaf through the phloem
(nutrient transport system) for systemic (long distance) infection
Í For most, but not all, plant viruses the extracellular form appears
to
be virus particles
Plant Viruses use Movement Proteins to Move Cell to Cell
without an Extracellular Phase and without Lysis

CELL 1 PD
CELL 2

MP

Move through plasmodesmata (PD) that have been altered

by a virus-encoded movement protein (MP)
Encapsidated virus particle (virion) not essential
Genome may be associated with MP or with CP
CP functions for insect transmission
 Classification of Viruses
[Lwoff, Horne, and Tournier (1962)]

Grouped by shared properties rather than the properties of

the cells or organisms they infect

Genome is primary criterion for classification

Characteristics used for classification

1. Nature of the nucleic acid in the virion (DNA or RNA)

2. Symmetry of viral capsid (icosahedral, helical, complex)

3. Presence or absence of an envelope

4. Dimensions of virion and capsid

Î Morphological distinctions belie similarities in genome organization

and replication strategy of different viruses
 No Matter How Complex, ALL Virions Have Their
Genome Encased in a Capsid

Animal RNA Viruses

Flint et al. Principles of Virology (2004)

 No Matter How Complex, ALL Virions Have Their
Genome Encased in a Capsid
Animal DNA Viruses

Flint et al. Principles of Virology (2004)

 Classification of Plant Viruses
Examples used in class:

Potyvirus: tobacco etch virus (TEV)

* *
Potexvirus: potato virus X (PVX)
Tobamovirus: tobacco mosaic virus
(TMV)
Tymovirus: turnip yellow mosaic virus
* *
Luteovirus: barley yellow dwarf virus
(BYDV)
* *
Polerovirus: potato leaf roll virus (PLRV)

Bromovirus: brome mosaic virus (BMV)

Tospovirus: tomato spotted wilt virus *
(TSWV)
Rhabdovirus: sonchus yellow net virus
(SYNV)
Caulimovirus: cauliflower mosaic virus
(CaMV)
Geminivirus: squash leaf curl (SqLCV), *
tomato golden mosaic (TGMV),
wheat dwarf (WDV) *
van Regenmortel et al. (1999) ICTV Report
 Baltimore Classification Scheme

Based on genetic system of each virus

Describes relationship between viral genome and its mRNA

mRNA = positive (+) sense

(also DNA of equivalent
polarity)

complement = negative (–) sense

Flint et al. Principles of Virology (2004)

 Infectious Subviral Agents

Prions (pathogens of animals)

ƒ Self-replicating proteins

Satellites (pathogens of animal and plants)

ƒ Encapsidated nucleic acid that depends on an unrelated
helper virus for its replication
ƒ If it encodes its coat protein, it is a satellite virus

Viroids (pathogens of plants)

ƒ Unencapsidated, highly structured small RNA (~240-390 nt)
Viroids are Highly Structured Small RNAs

From Agrios (1997) Plant Pathology 4th Ed.

 “Reverse Genetics”
+RNA genome DNA (cloned) introduce mutations
virions
infectious transcripts study individual
(cDNA) genes/proteins
vector
abrasion abrasion particle electroporation
(celite, bombardment [viruses, some]
carborundum)
agroinoculation

plants protoplasts

infectivity cell fractionation sections extract fix

live
symptoms (nuclei, chloroplasts,
membranes, soluble)
immunogold
immunogold EM viral proteins, EM
GFP
virions replication
EM
(blots) immune
viral proteins, immune fluorescent fluorescent
replication, mRNAs staining staining
(gels, blots)
GUS, GFP
reporter genes
 Virus Life Cycle: See a problem and figure out
how to surmount it
attachment/penetration*
Enters cells and multiplies,
usually at the expense of the
host cell
genome
uncoated Will use host cell machinery

(RNA = cytoplasm
mRNA replication DNA = nucleus) What are the molecular and
cellular events that allow virus
multiplication?
proteins
What steps are virus-specific
(potential targets for resistance)?

progeny virion assembly How do viruses modify host

cells to promote their own
multiplication?

“release” *
 RNA VIRUSES
ƒ ADVANTAGE
‹ RNA-dependent RNA polymerases do not require a primer
) replication can be initiated at the ends of a linear molecule
) 3 exceptions: picornaviruses [polio] (protein primer), influenza and
bunyaviruses (stolen caps)

ƒ PROBLEMS
‹ Cells do not express the enzymes required to initiate replication of viral
RNA
) virus must provide its own RNA-dependent RNA polymerase
(RdRP: replicase, transcriptase)
> Eukaryotic RNA viruses replicate in the cytoplasm
• exceptions: influenza, retroviruses
‹ Do not contain same controlling elements as the host to regulate gene
expression
) regulate gene expression in unique ways
) distinguish mRNAs from replication templates
‹ Viral genes organized and translated like host genes
) eukaryotic RNA viruses must reconcile genome structure with requirement for
monocistronic mRNAs
 Virus Life Cycle
entry/penetration*
Viral genomes are compact
•Strategiesto expand
genome translation their coding capacity
proteins and regulate their gene
uncoated
expression
replication

Rep •Transcriptional
transcription and translational
mRNA
•Multiple
proteins from
one gene (translation unit)
vRNA MP(s)
CP Viruses play by the rules

•Obey host rules for mRNA

virus particles translation
cell-to-cell
movement

transmission
 Reading

Flint et al. Principles of Virology (2004) 2nd Edition

ƒ Chapter 1 “Foundations of Virology”
ƒ Chapter 3 “Genomes”

Lazarowitz (2001) The Plant Viruses, pp 1-9.

Agrios (1997) Plant Pathology, Ch. 14 pp 479-508