VIROLOGY - MCB 5505

VIRUS FAMILY: PODOVIRIDAE (podo = foot) - PHAGE T7

I. DISTINGUISHING CHARACTERISTICS
A. MEDIUM-SIZED VIRUS WITH DS, LINEAR DNA
B. ICOSAHEDRAL WITH SMALL "FOOT" AT ONE VERTEX
C. MAKES ITS OWN RNA & DNA POLYMERASE
D. SHUTS DOWN HOST TRANSCRIPTION & DESTROYS HOST DNA
E. HAS TWO TYPES OF PROMOTERS, EARLY AND LATE
F. UNUSUAL DNA REPLICATION (RECOMBINATION EVENT)

II. STRUCTURE (for phage T7)

A. SIZE: 50 nm in dia., 45 mD
B. ENVELOPE: NO
C. NUCLEOCAPSID
1. NUCLEIC ACID
a. TYPE: DNA BALTIMORE TYPE: I
b. STRANDED: DS, LINEAR, TERM. REP.(160 bp)
c. POLARITY: +-
d. MOL. WT.: 26.6 mD; 39,936 N
e. # GENES: 55 ORFs (30 known genes)
2. GENETIC (PHYSICAL) MAP:

ppE.coli-------> pT7 -------------------------------------------->

=========================================/==/================
0.3 0.7 1 2 3 4 5 6 7 8 9 10 14 15 16 17 18 19

Gene 1 product is a T7 RNA polymerase. Genes 2-5 are involved in

DNA replication (and digestion of the E. coli genome). Genes 6-19
make head and tail virion proteins. Genes 0.7 & 2 code for
proteins that inactivate the E. coli RNA polymerase. Arrows
indicate transcription.
[There are many T7 promoters (pT7) between genes 1.3 and 19.]

3. CAPSID
a. SYMMETRY: ICOSAHEDRAL; T = 7, laevo
b. CAPSOMERS: 60 p10, 12 p10 ?
c. SIZE: 60 nm DIA; Tail is 20 X 10 nm
d. COMPOSITION - PROTEINS:

420 copies ? Major Coat - Head p10 35 kD

few copies Coat ? p10* 42 kD
few copies Head-Tail (portal) p8 60 kD
few copies Core p13 13 kD
few copies Core p14 15 kD
few copies Core p15 87 kD
few copies Core p16 150 kD
--- copies Tail p11 20 kD
--- copies Tail p12 93 kD
--- copies Tail Fiber p17 62 kD

VIRUS FAMILY: PODOVIRIDAE

III. CLASSIFICATION AND CHARACTERISTIC MEMBERS

GENERA PROPERTIES MEMBERS

T7-like Phage T7, T3, H, W31, R
φ29-like Phage φ29,
P22-like Phage P22,
Many unclassified

IV. VIRAL MULTIPLICATION

A. ABSORPTION. The virus (T7) binds to its host cell
via the “foot” that radiates from one of the
vertices. The 6 tail fibers attach to an E.coli
surface structure, probably on the OM.
B. PENETRATION. Only the DNA enters, with pilot
protein(s). The DNA enters slowly, always
beginning at the gene 0.3 end. One theory is that
transcription of the DNA slowly pulls the genome
into the cell.
C. UNCOATING. NA.
D. GENE EXPRESSION. The entering DNA has a series of
E. coli RNA polymerase-σ70 promoters in front of
the 0.3 gene. A transcription terminator is after
gene 1.3; thus, the early transcripts are made by E.
coli and include genes 0.3 --> 1.3. Gene 1 is the
T7 RNA polymerase, and genes 0.7 and 2 make
proteins which inactivate the E. coli RNA polymerase.
After a few minutes only T7 transcripts are made.
The early-late and late transcripts are from T7
promoters,and include DNA replication functions and
the phage virion proteins, respectively.
E. GENOME REPLICATION. A specific T7 DNA polymerase
(gene 5) is made, along with DNA endo- and exo-
nucleases (genes 3 & 6), a DNA binding protein
(gene 2.5), DNA ligase (gene 1.3), and primase
(gene 4). E. coli’s DNA is completely degraded and
nucleotides are used to make T7 DNA. The ori is
after gene 1, and DNA is replicated bidirectionally.
Concatemers are made from the new monomers by a
recombinational event involving the terminal repeats
of 160 bp.
F. ASSEMBLY. The virion consists of a head, short
tail (foot) and six thin tail fibers. The head is
composed primarily of one protein, p10- 420 copies,
and various internal proteins. Three tail protins
are present and a portal protein, p8. p18 & p19
are involved with final assembly and DNA cutting.
G. BUDDING AND/OR RELEASE. Gene 3.5 (a lysozyme) and
late gene 17.5(a holin)cause lysis at about 30 min.