VIROLOGY - MCB 5505

VIRUS FAMILY: MICROVIRIDAE (micro=small, minute)

I. DISTINGUISHING CHARACTERISTICS
A. Small, single-stranded, circular DNA phage
B. Model for circular genomes & plasmids
C. Rolling circle DNA replication
D. φX174 first DNA genome sequenced (Sanger et al, 1977)
E. Extensive overlapping genes (Barrell, 1976)
II. STRUCTURE (for φX)
A. SIZE: 35 nm in diameter, PW=6.4MD, 114S
B. ENVELOPE: NONE
1. GLYCOPROTEINS:
2. OTHER PROTEINS:
3. MATRIX PROTEIN:
C. NUCLEOCAPSID
1. NUCLEIC ACID
a. TYPE: DNA BALTIMORE TYPE: II
b. STRANDED: SS, CIRCULAR
c. POLARITY: +
d. MOL. WT.: 1.7 MD
e. # GENES: 11
2. GENETIC (PHYSICAL) MAP: φX174
5386 N - CIRCULAR
A*
1 |------| 5386/1
-|-------------|-----|--------|---|-|--------|-|------|------|-
A C D J F G H
|---|--| |---|
B K E
PaÆ PbÆ PdÆ

3. CAPSID
a. SYMMETRY: Icosahedral, T=1
b. CAPSOMERS: 12 pentons (5 pF, 5 pG, 1 pH)
c. SIZE: 27 nm
d. COMPOSITION
(1) PROTEINS
60 copies Major capsid (F) protein, 60 kD
60 copies Major Spike (G) protein, 20 kD
12 copies Spike/Pilot (H) protein, 37 kD
(2) OTHER COMPONENTS
60 copies Internal (J) protein, 4 kD
(DNA binding)
4. NON STRUCTURAL PROTEINS
Nickase/ligase (A) protein 60 kD
Host DNA turnoff (A*) protein 37 kD
Scaffolding (in) (B) protein 30 kD
Maturation (C) protein 6 kD
Scaffolding (out) (D) protein 14 kD
Lysis (E) protein 10 kD
Membrane (K) protein 8 kD
VIRUS FAMILY: MICROVIRIDAE
III. CLASSIFICATION AND CHARACTERISTIC MEMBERS

GENERA PROPERTIES MEMBERS

Microvirus Coliphage φX174, G4, α3
Bdellomicrovirus Bdellophage MAC1
Chlamydiamicrovirus Chlamydiaphage Chp-1
Spiromicrovirus Spiroplasma phage SpV4

IV. VIRAL MULTIPLICATION

A. ABSORPTION: Spike or pilot protein attaches to
specific lipoproteins on surface of E. coli.

B. PENETRATION: Pilot protein enters with DNA.

C. UNCOATING: During penetration.

D. GENE EXPRESSION: The ss, circular DNA genome is
converted to a ds DNA, the RF or Replicative Form.
E. coli enzymes are responsible. Starts between F
& G. Transcription is initiated from several
(three) promoters. Proteins are made in different
amounts depending on the strength of promoter; mRNAs
are polygenic. Circular bidirectional replication
proceeds from φX origin (in A gene), pA, a nicking
protein is required for this. Later rolling circle
replication also proceeds from this site, with pA
remaining attached to the 5’-end of single stand.
Concatemers are formed. pA nicks and recloses ss
circles - new genomes.

E. GENOME REPLICATION: ABOVE - What are RFI, RFII, and

RFIII?

F. ASSEMBLY: The capsid self assembles using the

scaffolding protein (pB) which provides a temporary
core. An external scaffold is provided by pD.
Several precursor particles appear before the entire
capsid assembles: 6S, 9S, 12S, 108S & 132S. The
mature particle with DNA is 114S.

G. BUDDING AND/OR RELEASE: Particles accumulate in

the cell, and if no lysis occurs or is late, more
particles accumulate. Lysis is catalyzed by
hydrophobic protein, pE.