Micro exam 5

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What are some general characteristics of viruses

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1

What are some general characteristics of viruses

Minuscule, acellular, infectious agents having either DNA or RNA

cause infections of humans, animals, plants, and bacteria

Cannot carry out any metabolic pathway

cannot reproduce independently

have no cytoplasmic membrane, cytosol, organelles

has an extracellular (ex. virion) or intracellular state (ex. genome)

Neither grow nor respond to the environment

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Extracellular state of viruses

called a virion

has a protein coat (capsid) surrounding nucleic acid

Some have envelopes

outermost layer (capsid or envelope) provides protection and recognition sites for host cells

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Nucleocapsid

nucleic acid and capsid together

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4

Envelope

nucleocapsid surrounded by envelope of host cell’s membrane

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Intracellular state

capsid removed

virus exists as nucleic acid

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Virulent (lytic) infection

replicates an destroys host

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lysogenic infection

host cell genetically altered because viral genome becomes part of host genome

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Single-stranded DNA or RNA of viruses may be plus sense ______ or minus sense _____

same as mRNA

complementary to mRNA

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Genetic material of viruses

show more variety in nature of their genomes than do cells

primary way that scientists categorize and classify viruses

may be linear and segmented or single and circular

much smaller than genomes of cells

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10

Most viruses infect only particular host cells why

due to affinity of viral surface proteins for complementary proteins on host cell surface

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Host viruses may be so specific they only infect particular kind of cell in a particular host, what is an example of this

HIV

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Generalists infect many kinds of cells or many different hosts, what is an example of this

West Nile Virus

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All types of organisms are susceptible to what

Viruses

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14

Bacterial viruses infect

bacteriophages (model systems for viral infections)

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Archaeal viruses attack

Archaea

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Animal viruses attack

animals (this is extensively studied)

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plant viruses attack

plants

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bacteriophage

a virus that infects and replicates within a bacteria

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capsid

provides protection for viral nucleic acid

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Capsids help the virus do what

attach to the host cell

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Capsomeres are what

proteinaceous subunits that make up capsids

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Capsomere

individual protein molecules arranged in a precise and highly repetitive pattern around the nucleic acid making up the capsid

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what are the three basic shapes of viruses

Helical, polyhedral, and complex

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Helical shaped

capsomeres are bound together forming a circular tube around nucleic acid

rod shaped viruses have helical symmetry

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Polyhedral shape

Spherical shape (a geodesic dome)

Icosahedron has 20 sides

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complex shape

capsids can have many different shapes

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Helical symmetry

rod-shaped viruses (ex. tobacco mosaic virus or TMV)

length of virus determined by length of nucleic acid

width of virus determined by size and packaging of capsomeres

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Icosahedral symmetry

spherical viruses (ex. human papillomavirus)

most efficient arrangement of subunits in a closed shell

requires the fewest capsomeres

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bacteriophages have what 2 parts

the head and tail

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Viral envelope

acquired from host cell during viral replication or release (envelops is portion of membrane system of the host)

composed of phospholipid bilayer and proteins (some proteins are virally coded glycoproteins - spikes)

Envelope proteins and glycoproteins often play role in host recognition and for release after infection

Enveloped viruses are more fragile than naked viruses

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viral classification is based on

type of nucleic acid

presence of an envelope

shape

size

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32

viral genera have only been organized into what, why?

families

the relationship between viruses is not well understood by taxonomists

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Viral replication is dependent on what

hosts’ organelles and enzymes to produce new virions

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lytic replication

viral replication usually results in death and lysis of host cell

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what are the 5 stages of the lytic replication cycle

attachment

entry (penetration)

synthesis

assembly

release

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36

The most complex penetration mechanisms are found in what type of virus

tailed bacteriophages

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37

Virions attach to cells via tail fibers that interact with what

polysaccharides on Escherichia coli LPS layer

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38

Describe the attachment and entry of a bacteriophage

Virions attach to cell via tail fibers that interact with polysaccharides on the LPS layer of Escherichia coli

tail fibers retract, and tail pins contact the cell wall

T4 lysozyme forms small pore in peptidoglycan

tail sheath contracts, and viral DNA passes into the cytoplasm

capsid stays outside

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39

Describe the assembly process of bacteriophages

Base → tail → sheath → DNA → Capsid → mature head → tail fibers → mature virion

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Circular permutation

feature of many virus genomes where same genes are arranged in different orders

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terminally redundant

some DNA sequences duplicated on both ends

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T4 encodes what

primases, helicases, and 8-protein DNA replisome complex

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Small DNA viruses use what for genome replication

host cell’s DNA polymeras

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Complex DNA viruses use what for genome replication

their own polymerases (ex. T4)

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T4 first replicates as a unit then forms what

concatemer (several genomic units recombined)

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Virion synthesis takes how long

~ 30 minutes and ends in release

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Early proteins

enzymes needed for DNA replication and transcription

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Middle and late proteins

head and tail proteins and enzymes required to liberate mature phage particles

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How does the genome get into the virion head

the genome is pumped into the head under pressure using ATP

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What are the three stages of packaging the T4 genome into virion head

Proheads (bacteriophage head precursors) assembled

Packaging motor assembled at opening

Double-stranded linear genome pumped into prohead using ATP

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51

Virulent mode

viruses lyse host cells after infection

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Temperate mode

viruses replicate their genomes in tandem with host genome and without killing host

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what are the two viral life cycles

virulent and temperate

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virulent bacteriophages were the first viruses studied in detail what have we learned

Bacteriophages contain linear, dsDNA genomes that infect and kill their host

the study of these viruses established many fundamental principles of molecular biology and genetics

Examples include; T1, T2, T3, T4… T7

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Temperate viruses

can undergo a lytic (short term) or lysogenic life cycle (long term)

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Lysogeny

state where most virus genes are not expressed and virus genome (prophage) is replicated in synchrony with host chromosome

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lysogen

host cell that harbors temperate virus

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lysogeny is maintained by phage-encoded

repressor proteins

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What are the general steps in a lytic pathway starting with a temperate virus

temperate virus attaches to the host cell

injects the viral DNA

Lytic events are initiated

Phage components are synthesized and virions are assembled

lysis of the host cell and release of new phage virions

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60

What is the general pathway of temperate virus through a lysogenic pathway

Temperate virus attaches to host cell

Injects viral DNA

Viral DNA is integrated into host DNA

(cell becomes a prophage)

Viral DNA is replicated with host DNA at cell division

(an induction step can result in these cells switching over to a lytic pathway where host cell is lysed)

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Bacteriophage lambda infects what

E. coli

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Bacteriophage lambda structure

linear, dsDNA virus with head and tail

complementary, single-stranded “cohesive” regions have 12 nucleotides long at the 5’ terminus of each strand

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When bacteriophage lambda penetrate what happens in the host cell

DNA ends base pair, forming the cos site, and DNA ligates and forms double-stranded circle

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when lambda enters the lytic pathway what happens

lambda synthesizes long, linear concatemers of DNA by rolling circle replication

genome-sized lengths cut at cos sites; genomes packaged into phage heads

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When lambda is lysogenic its DNA integrates into E. coli chromosome where

at the lambda attachment site

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what does the lambda use to attach at the attachment site

lambda integrase

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what induces the lytic pathway of bacteriophage lambda

cell stress

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How is the lysogenic DNA added to the host cell DNA

a site-specific endonuclease creates staggered ends of phage and host DNA

Lambda genome integrates and gaps are closed by DNA ligase

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69

What are the key elements of regulation

two repressor proteins:

cl protein (lambda repressor) and cro repressor

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when do repressors accumulate

after infection

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cl protein (the lambda repressor)

causes repression of lambda lytic events

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cro repressor

controls activation of lytic events

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what controls the infection outcome

first repressor

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When cro is not repressed and lambda low which event happens

cell lysis

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when cro is repressed and lambda high which cycle does it go through

lysogeny

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76

animal virus infection

major tenets (capsid and DNA/RNA genome, infection and takeover of host, assembly, and release) are universal

classified by genomes

most human viral diseases are caused by RNA viruses

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What are the two key differences between animal virus infections

Entire virion enters the animal cell

or

Eukaryotic cells contain a nucleus, the site of replication for many animal viruses

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Unlike prokaryotes, how does a virion enter the animal cell

the entire virion enters the cell, not just the genome

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ssRNA are

large

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dsDNA are

HUGE

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overview of viral infection of animal cells

bind specific host cell receptors, typically used for cell-cell contact or immune function

different tissues and organs express different cell surface proteins (often viruses only infect certain tissues)

Entry usually occurs by fusion with cytoplasmic membrane or endocytosis

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82

what are 3 general steps of virion entry via cell recognition and membrane fusion/endocytosis

Rotavirus virion is bound to cell surface proteins/receptors

virion is engulfed by host cell membrane

compartmentalization

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83

Viral DNA-genomes replicate where

in the nucleus

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84

Viral RNA genomes replicate where

most viral RNA is replicated or converted to DNA within nucleocapsid (not in the nucleus, just in the cytoplasm)

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where does uncoating or shedding of the capsid occur

at the cytoplasmic membrane or cytoplasm

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what are examples of positive-sense ssRNA virus

polio and coronavirus

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what are examples of negative-sense ssRNA virus

rabies and flu

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Double-stranded RNA virus examples

rotavirus

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What is the process of protein synthesis and genome replication in animal RNA viruses with a positive-sense ssRNA virus

Receptors on cytoplasmic membrane of host recognize the +ssRNA virus

+ssRNA is used in translation of viral proteins; genome acts as mRNA

transcription by viral RNA polymerase using a -ssRNA complementary strand as a template

Transcription occurs to copy +ssRNA

Assembly of proteins and genome

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90

what is the process of protein synthesis and genome replication in animal RNA viruses with a negative-sense RNA virus

Receptors on cytoplasmic membrane of host cell recognizes -ssRNA virus

-ssRNA virus is inserted into the host cell

Transcription by RNA-dependent RNA transcriptase

Complementary +ssRNA to act as a template and mRNA

+ssRNA can be translated into viral proteins

+ssRNA can be further transcribed into copies of -ssRNA

Copies of -ssRNA and viral proteins can be assembled for dispersal

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91

what is the process of protein synthesis and genome replication in animal RNA viruses with a double-stranded RNA virus

receptors on cytoplasmic membrane of host recognise dsRNA virus

dsRNA in inserted into the cell

Unwinding of dsRNA yields -ssRNA and +ssRNA

-ssRNA is transcribed by viral RNA polymerase to make complementary RNA strands

+ssRNA acts as template and as mRNA

Translation of viral proteins from +ssRNA

Viral proteins and viral genome (dsRNA) are assembled into new viruses

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92

Most DNA viruses assemble in the

nucleusmost

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Most RNA viruses develop where

solely in the cytoplasm (anywhere outside the nucleus)

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The number of viruses produced depends on what

on the type of virus and size and initial health of the host cell

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95

Enveloped viruses can cause what

persistent infections (doesn’t kill the host cell immediately) via budding

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Naked viruses are released by what

exocytosis or lysis

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What is the process of budding

Viral capsid is drawn to the plasma membra

Budding of the virus occurs, encapsuling the virus

cytoplasmic membrane of host envelopes the virion

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Virulent infection

lysis of host cell, most common (especially naked cells)

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Latent infection

viral DNA exists in host genome and virions are not produced; host cell is unharmed unless/until virulence is triggered

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Persistent infections

release of virions from host cell by slow-budding does not result in cell lysis (infected cell remains alive and continues to produce virus)

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