Problems viruses must solve

SV40

Attachment and entry:

receptor?
stimulate endocytosis or fusion?

Uncoating:

trigger? host or viral function?

Genome replication:

host or viral enzymes?

nuclear or cytoplasmic?
end problem?

Protein synthesis:

multiple proteins, minimal genome

use host machinery effectively
regulation, timing
block host functions?

Part of papovavirus family

small (45 nm)
naked, icosahedral
dsDNA

Polyomavirus genus
SV40
mouse Py
human BKV
human JCV

Not associated with disease

human polyomaviruses cause disease if immunocompromised

Maturation:

structure
genome packaging

Isolated in 1960

Release:

lysis how?

Used extensively in studies of DNA replication

contaminant of Vero cells used to grow OPV virus

SV40 structure

SV40 structure

Icosahedron:

Icosahedron

Hollow sphere (approximately) of triangular faces

Simplest: 20 triangular faces
60 protein subunits minimum
One protein minimum
To enlarge the sphere, triangulate the triangular faces
T = triangulation number = number of triangles in a face

T=1

T=3

SV40 replication
Attachment
Receptor appears to be MHC I
VP1 (only exposed protein) binds
receptor

Entry
Receptor-mediated endocytosis
VP2, VP3 may interact with
membrane
Probably enters ER
Transported into nucleus;
mechanism uncertain
Uncoating occurs in nucleus;
requires VP2, VP3

T=7
3 capsid proteins: VP1, VP2, VP3
60 subunits with 5 VP1 proteins each

T=4

SV40 replication
Early gene expression
Genome is minichromosome with
histones (5243 bp)
Host factors recognize
TATA box and enhancer
of early promoter (PE)

SV40 replication
Early gene expression
small-t and large-T antigen mRNA made by alternative splicing
Translation by host factors

SV40 replication
Late gene expression
T Ag binds promoter region
Activates PL while blocking PE

VP1 and VP2/3 made from

two different mRNAs (alt.
splicing)
VP3 made by ribosome
skipping on VP2 mRNA
Agno protein made from
mRNAs with different start
site

SV40 replication
Genome replication
T Ag binds host Rb and p53
Removes contact inhibition
DNA replication initiated
Blocks induction of apoptosis
T Ag binds SV40 origin acts as initiator protein
Host helicase, primase, polymerase, etc. replicate DNA
Circular DNA: no ends, so no end problem

SV40 replication
Assembly & Release
Virus proteins have
nuclear localization
signals
Assembly occurs in
nucleus
Some virus transported
to surface in vesicles
Cell eventually lyses
(virus production
relatively slow)

SV40 transformation
Abortive infection in nonpermissive cells (e.g., mouse)
Entry and T Ag synthesis occur normally
No DNA synthesis
DNAP cant interact with T Ag at origin?
T Ag binds Rb and p53
Cells lose contact inhibition and replicate: transformed
Rarely, SV40 DNA integrates into nuclear DNA
Stable transformation if this occurs