J. gen. Virol.

(I975), 27, t35-149 Printed in Great Britain

I35

A Classification of Virus Groups Based on the Size of the Particle in Relation to Genome Size
By R. E. F. M A T T H E W S

Department of Cell Biology, University of Auckland, Auckland, New Zealand (Accepted 6 January I975)
SUMMARY

For 59 different viruses, when the amount of nucleic acid in the particle is related either to the dry weight of the particle or to the particle volume, two classes of virus groups emerge - those with enveloped or those with geometrical particles. The enveloped viruses have particles with the following properties: (i) about 4o Io n daltons of anhydrous weight per Io n daltons of nucleic acid; (ii) a particle volume of about 2 io 5 nm 3 per I o daltons of nucleic acid; (iii) a limiting lipoprotein membrane. These properties are qualitatively and quantitatively close to those of prokaryotic cells. The geometric viruses have particles with roughly one-tenth the anhydrous mass per unit of nucleic acid and one twenty-fifth the particle volume per unit of nucleic acid. They do not possess a limiting lipoprotein membrane.

INTRODUCTION

In the various classifications of viruses that have been proposed the choice of characters in the hierarchy has been quite arbitrary, and, among the characters used to make the major subdivisions, there has been no natural reinforcement of one property by another. Where virus size has been considered as a parameter, only linear dimensions have been used. Thus Lwoff and his colleagues used the possession of D N A or RNA for the first division in their classification; capsid symmetry for the second; and the presence or absence of an envelope for the third (Lwoff, H o r n e & Tournier, I962; Lwoff & Tournier, I97I ). Melnick 0973) used the same divisions for his most recent classification of animal viruses. The diameter of the virus particle and the mol. wt. of the nucleic acid were the 7th and 8th parameters used by this author. Davis et al. 0973) in their table listing characteristics of viruses use the symmetry of the capsid for the first division; then the presence or absence of an envelope; then the presence of D N A or RNA. In the first report of the International Committee on Classification and Nomenclature of Viruses (Wildy, I97I) the 43 virus groups described are arranged according to the nucleic acid content of their particles. The cryptogram (a summary of 'important' virus features) contains no information on membranes or lipid content. Indeed the cryptogram subcommittee dismissed a suggestion that the presence or absence of lipid should be included (Wildy, I970. In this paper I wish to show that when the mol. wt. of nucleic acid in the particle is related either to the dry weight of the particle or to the particle volume (nm a) two classes of virus groups emerge. One of these classes possesses a lipoprotein envelope. The other does not.

136

R.E.r.

MATTHEWS

Table I. Data for the groups of enveloped viruses and some prokaryotes
Nucleic acid mol. wt. ( lO-6) Particle mass (dry) ( x io -e) 1"5 IO5 1.6 lO4 2"4 x 1o4 I.I lO4 3200 2700 Particle dimensions (nm)
-

Virus group or cell

Particle volume (nm 3)

lipid by wt. 7 22 IO-ZO

References McQuillen (I965) Loewy & Siekevitz (1969) Loewy & Siekevitz (I969); Davis et al. 0973) Moulder (I964) McAuslan (I969) Davis et aL (1973); Wagner et al. (1974); Russell, Watson & Wildy (1963) Wildy (1971); Scholtissek, Drzeniek & Rott (1969) Harvey (1973); Bellamy, Gillies & Harvey (1974); Quigley, Rifkin & Reich (I971) Howatson (197o); Wildy (1971); Knudson (1973) Wildy (i97i); Harrison et al. (I971); Pfefferkorn & Hunter (1963) Wildy (197I) Green (1969)

Escherichia eoli 5000 Dialisterpneumosintes 750

lO9 500 500 IO00 I'95 I08 5oo 6'5 lo 7

Mycoplasma Psittacosis group Poxvirus (vaccinia) Herpes virus (herpes simplex)

5o0 400 16o 82

450 270 270 I40 18o

4"7 x io 7 I0 106 3"0 x lO6

47 5"6 22

Paramyxovirus (NDV)

660

x50

1"7 IO~

27

Leukovirus Rous (dry) Rous (hydrated)

4"2 4"2

294 294

90 I5O

3"8 Io 5 I '76 X IOn

- 3I~ t

Rhabdovirus (VSV)

3"5

I7O

17o x 75 diam.

7"4 x I o6

2o

Alpha virus (Sindbis)

3"0

53

1.8 lO5

26

Flavi virus (Dengue type I) Myxovirus (influenza)

3"0 3"0

43 250

50 Ioo

6"5x lO4 5"2 x to e

Present 18"5

METHODS

Volume calculations. The volumes of all the particles required for infectivity were added together when calculating the volume of multiparticle viruses and the total amount of nucleic acid required for infectivity was used to calculate mol. wt. of nucleic acid. For viruses with tails, total volume was estimated. Groups of viruses considered. Wildy 0971) lists 43 virus groups. There are sufficient data for calculations to be made for 4o of these. The data for these groups and 19 additional groups are given in Tables I to 4. The definition of virus groups is sometimes quite arbitrary in the present state of knowledge. Most of the viruses considered are the best-studied examples of big groups of viruses whose members share all the properties being discussed (e.g. one rhabdovirus, VSV). However, reovirus and two other double-stranded R N A viruses are included. Two rather different iridescent viruses have also been listed.

A classification using virus and genome size

RESULTS

137

Two classes of viruses When the dry mass of the virus particles (in daltons) is plotted against mol. wt. of their nucleic acid (in daltons) two classes e m e r g e - those bounded by a lipoprotein membrane and those without such a membrane (Fig. I). The same two classes are apparent when the volume of the virus particles (nm 3) is plotted against the mol. wt. of their nucleic acid (Fig. 2). The lines in Fig, r and 2 were drawn by inspection. They have a slope of I-0 and are intended to assist in identifying the two classes of viruses. I shall use the term enveloped for those viruses which possess an external lipoprotein bilayer membrane which is necessary for the integrity of the particle. As noted below, certain particles containing lipid are excluded from this class. Enveloped virus particles have a particle dry mass of roughly 40 I06[I06 daltons of nucleic acid, and a particle volume of approx. 2 lO5 nmZ/Io 6 daltons of nucleic acid. The four prokaryotic cells shown in Fig. I and 2 have similar values for these parameters. In contrast, particles of viruses without a limiting lipoprotein envelope (geometrical viruses) have roughly one-tenth the particle dry mass, and one twenty-fifth the volume, per unit of genetic material possessed by the enveloped viruses and the prokaryotic cells. There is probably substantial error in the estimates of volume for some of the enveloped viruses since estimates have been made on dried preparations. The extent of this error is indicated by recent data for hydrated RSV (Table I, Fig. 2). It is possible that when diam. in solution have been estimated for the alpha, flavi and herpes viruses, that similar corrections may be made that will bring these viruses more closely into line with others of the enveloped class. The volumes for the geometrical viruses are also based on measurements made in the dry state. These may also be substantially in error, at least for the larger viruses that possess an outer layer of protein subunits surrounding an inner core. The possible magnitude of such error may be judged from the data for reovirus, where we have an accurate determination of the diam. in aqueous solution (Harvey, ~973) (Fig. 2). Inspection of Fig. I and 2 shows that the enveloped virus particles fall into two subclasses: (a) those with double-stranded D N A and (b) those with single-stranded RNA. Fig. I and z also show that the geometrical viruses fall naturally into three subclasses: (i) all the virus particles that contain double-stranded nucleic acid (DNA or RNA) have a nucleic acid content/> 3"4 lO6 daltons; (ii) all the virus particles that contain singlestranded nucleic acid (DNA or RNA) have a nucleic acid content ~< 4"1 lO6 daltons; (iii) the viruses with rod-shaped particles that cluster near the single-stranded icosahedral viruses. These viruses tend to have a high volume or particle mass per unit of nucleic acid because of the less efficient packing of nucleic acid by protein subunits in a helical array, compared with icosahedrat. This can be illustrated by comparing TMV and TYMV. Both have single-stranded R N A with a mol. wt. of about z.o x lO6. The coat protein subunit of TMV has a mol. wt. of 17 5oo, while that of TYMV has a tool. wt. of 2o ooo. TMV has a dry particle mass of 39.o Io 6. For TYMV the dry mass is 5"4 Io6. TYMV RNA, reconstituted in vitro with TMV protein produces a rod with the dimensions of TMV (Matthews, 1966).

x38

R.E.F.

MATTHEWS

T a b l e 2. D a t a f o r the groups o f geometrical viruses containing

double-stranded nucleic acid

Mol.
Wt.

of Particle nucleic dry acids mass Virus g r o u p Iridovirus R M I V (mosquito) T2

( 10 -6) ( 10 -6)

Particle dimensions (nm)

215"

Particle volume ( n m 3) 5'15 106

Lipid (~o by weight)

References

464 I3O

2750 325

3"9 W a g n e r et al. (I973) o M a t t h e w s (I969)

~5CbK

130

230

F r o g virus 3

I3O

430

Head 34"6 x lO4 95 65 diam., tail I 0 0 20 diam. Head _~ 6.2 x io ~ 195 62, tail 275 long 135 1"28 lO6

A g a b i a n - K e s h i s h i a n & Shapiro (197o)

14

Iridovirus (Tipula)

IO5

55I

13o

I ' I 4 x lO 6

Baculo (nuclear polyhedrosis

8o

-~ 65o

4oo 7o

1.5 x lO 6

L u n g e r & C a m e 0 9 6 6 ) ; Smith & M c A u s l a n (I969); T a n & M c A u s l a n (197I) 9 K a l m a k o f f & T r e m a i n e (I968); Kelly & A v e r y (1974); Kelly & Vance (I973) o-2 Bergold & Wellington (1954); Wildy (I971); 13ellett et al. (1973) o O o Bayer & B o k a r o v ( 1 9 7 3 ) ; M a t t h e w s (1969) B r o w n (1972) A d a m s (1959; D a v i s o n & Frieii~lder (1962); Studier (1972). Barrett et al. (I973)

Bombyx)
Lambda L-PPI T7 33 27 25 66 68 130 H e a d 64, tail 165 x 13"6 59 H e a d 47, tail
I n 15 P2 22 --

16-I lO5 1"O7 x IO5 5"5 x ~o4

Adenovirus PM2 Reovirus (dry) Reovirus (hydrated) Wound turnout virus Insect cytoplasmic polyhedrosis Bacteriophage ~29

22 I8 15 15 15 15
II

183 129 130 I3O 7I 54

18

H e a d 58, tail 135x I7 7o 6o 75 98 70 65 Head 32 3I, tail 32 6 H e a d 44, tail 135 17 53 50

I. 3 x io ~

I7-8 IO4 I'13 x io 5 2'2 IO 5 4"9 X 106 1.8 x to ~ 1-43 ro~ 2"5 x 1 0

o io 0t o( o o o

Pifia & G r e e n (1965); G r e e n et al. i967); Wildy (i971) Espejo & Canelo (I968) H a r v e y (I973); W o o d (I973); Farrell, H a r v e y &]3ellamy (1974) W o o d (1973) W o o d (1973) A n d e r s o n , H i c k m a n n & Reilly (1966); R u b i o et al. (1974) Barrett et al. (I973) Wildy (I97I); G r e e n (1969) Shepherd (I97O); Shepherd, B r u e n i n g & W a k e m a n (197o); Bellett et aL (1973) Wildy (I971)

P4 Papillomavirus (Shope rabbit) Caulimovirus

6"7 5 4"5

-4o 28

7"5 x lO4 7'7 lO4 6"5 IO4

o o o

Polyoma

3"4

26

43

4 ' 1 2 104

* M e a n value between t h o s e for stained a n d thin section preparations.

A classification using virus and genome size

Table 3- Data for the groups of geometrical viruses containing

I39

single-stranded nucleic acid

Nucleic acid Particle mol. wt. drymass ( x i o -6) ( x 1o -6) 4-t 3"7 3"5 3"4 3.0 ~ 3.0 2"9 2.8 2.8 2.6 2.6 2.6 2-6 2'2 2.o 2"o 2.0 1"7 1.62 1"6 1% 1-4
I-1 1.1

Virus group CpMV NEPO BBWV CMV AMV CaI~ci PEMV

Particle dimensions (nm) 28 ( x 2) 27.5 ( x 2)

27(2) 28 (X 3) 58X 18;

Particle volume (nm ~) 2'3 x 1o4 2.3 X 10*

2 ' I 2 X IO4 3-5X 104 3.19X i 0 4

Lipid ( % by weight) o 0
0
0

References Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973); Lot et al. (I974) Bos & Jaspars (1971) Brown & Hull (1973) Hull & Lane (1973) Fulton (1971); Ghabrial & Lister (1974) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973); Bellett et al. (1973) Tinsley & Longworth (I973); J. F. Longworth (personal communication) Brown & Hull (1973) Matthews (1969); Spencer et al. (I972); Wildy (197I) Tinsley & Longworth (I 973); Wildy (I97 I) Brown & Hull (1973) Brown & Hull (1973) Brown & Hull (1973) Brown &I-lull (1973) Brown & Hull (1973) Kassanis (I97o)

t3'7 lO'6 11'5 5"5 18.2 7'0 I1'7 t8.1 I4'o 8"5 8"5 8"3 8"4 8"4 6-7 5'4 5"4 6.8 4"7 9"3 7"0 6'3
3"6 4"2

TSV (assuming 3 largest RNAs are needed) BMV Entero Cardio Equine rhino Foot and mouth H u m a n rhino Acute bee paralysis Parvovirus

49 I8; 38 x 18 3S 29 and 27 approx. 3 particles largest 28 26 ( x 3)

28 28 28 24 28 29

2'24 X 1 0 4 2"3I I04 2"8 x lO4 2'76 x I04

1"15 X 104 I'15 X 10 4 I ' 1 5 X 104 o,72 X 104 I'15 ~ 104 I'27 % 104

o o

o o o o o o o o o o o o o o 0 o o
o

22 28 25 2I 30 26 28"5
25 25

o.55 X lO4 I"15 lo 4 o,81 >~1o4 0'48 X IO4 1'4 104 o,92 x IO4 1,2 x I0 i
0'81 g I04 0.81 ~< 104

(Galleria)
TYMV X174 Latent rat TBSV TNV SBMV R17

Qfl

Satellite

0-4

2.0

17

2,6x lO8

T a b l e 4. Data f o r the groups o f rod-shaped viruses Nucleic Particle acid mass mass (dry) ( 1o6) ( IO-6) 3'8 3"6 3"6 2"1
2-o

Virus group Barley stripe mosaic virus Carla viruses TRV Potato X virus TMV fd virus

Particle dimensions (nm) 3 8 7 x 2 o ( 3) 65ox 12 305 x 25 515 x 13

3oox I8

Particle volume (nm") I2X IO4 7 x lO4 15 x lO4 7 x 104

7 - 6 x IO 4

References Lane (1974) Wetter (197 I) Harrison (197o) Bercks (197o) Klug & Caspar (196o) Frank & Day (x976); Ray (1968); Matthews (1969)

95 60 79 35
39

1"9

I6

88OX5

1'7 x I04

I40
I

R. E. F. MATTHEWS
I
I

E. coli
1011

/
Mycoplasma ( 7 ~

10~

PsittaDialister

109
O

Paramyxo
Rous

/
_ _

Rhabd~/ 108

.X_X/

/;

Alpha

, X xx

Flavi X l~Ik~x /
..{
I0? . .,,"

I I I 107 10s 109 101 Mol. wt. of nucleic acid Fig. I. Relationship between mol. wt. of nucleic acid and dry mol. wt. of particle for 59 groups of viruses and four prokaryotic cells. Q, cells and enveloped virus particles. The names of these are indicated, x, unenveloped virus particles containing double-stranded nucleic acid. I , viruses with unenveloped rod-shaped particles. 0, viruses with unenveloped icosahedral particles containing single-stranded nucleic acid.

106 105

I 106

Some other properties of the enveloped and geometrical classes of viruses Presence of lipid
All the enveloped virus particles contain lipid, usually 20 to 30 % by weight (Table I). None of the geometrical virus particles containing single-stranded nucleic acid have been reported to contain lipid. Five of the geometrical viruses have been shown to contain some lipid (Table 2). However, in these, the lipid does not form an outer bilayer membrane. It is present as a smaller proportion of the dry weight of the particle than found for the enveloped viruses (except poxvirus). Where it has been studied the composition of the lipid in the geometrical viruses differs significantly from the host lipid and much of the lipid is synthesized during virus maturation (e.g. Tsukagoshi & Franklin, I974). Frog virus 3 (FV3) and similar viruses have very large icosahedral particles which appear

A classification using virus and genome size

109
I I I I E. coli

I4I

Mycoplasma 108
Dialist

Psittacosis

107 S

106

Myxo . . ~

Rhabdo "ha~~Rous

I (D)

/ x

/Alpha(~
lOs Flavi I ( ~ xx
.e

" ~x

104

/
I

,I

I,,

lOs

lO6

107

108

109

10t

Mol. wt. of nucleic acid Fig. 2. Relationship between tool. wt. of nucleic acid and volume of particle (nm3) for 59 groups of viruses and four prokaryotic cells. G, ceils and enveloped virus particles. The two circles joined by a vertical line are for Rous hydrated (upper) and Rous dry (lower). x, unenveloped virus particles containing double-stranded nucleic acid. The two x joined by a vertical line are for reovirus hydrated (upper) and reovirus dry (lower). I , viruses with unenveloped rod-shaped particles. O, viruses with unenveloped icosahedral particles containing single-stranded nucleic acid. to acquire an envelope derived by budding from the plasma membrane (Darlington, Granoff & Breeze, I966). Nevertheless particles purified from disrupted infected cells are not enveloped but are infectious (Hauts, Gravell & Granoff, I974). Thus, the membrane is not an essential part of the virus particle. On other properties they fit well in the non-membrane bound class. The 14 ~ lipid found in unenveloped FV3 particles is probably an integral part of the virus as it is in PM2 (Harrison, Caspar, Camerini-Otero & Franklin, ~971 ; Willis & Granoff, I974). The baculoviruses are frequently referred to as having a membrane, but this appears to consist of a stacked disc array of protein subunits (Bellett, Fenner & Gibbs, I973) and

I42

R. E. F. MATTHEWS Table 5. S o m e e n z y m e s f o u n d in viruses Virus or group A. Enveloped groups Enzymes DNA-dependent RNA polymerase Nucleotide phosphohydrolase Protein kinase Endonuclease; Exonuclease Terminal riboadenylate transferase Protein kinase RNA-dependent RNA polymerase Protein kinase RNA-dependent DNA polymerase DNA ligase Exonuclease RNA-dependent RNA polymerase Protein kinase RNA-dependent RNA polymerase B. Geometrical groups RNA polymerase ATPase; lysozyme RNA-dependent RNA polymerase Nucleoside triphosphatases Two endodeoxyribonucleases Endoribonuclease Nucleotide phosphohydrolase Adenosine triphosphate phosphohydrolase Endonuclease (DNA) Endonuclease References* Kates & McAuslan (1967) Gold & Dales (I968) Kleinman & Moss (I973) Aubertin & McAuslan (I972) Brown, Dorson & Bollum (1973) Randall et al. 0972) Huang, Baltimore & Bratt (I97I) Roux & Kolakowski 0974) Mitzutani & Temin 0970 Mitzutani et al. (I970 Chang et at, (I974) Imblum & Wagner (2974) Bishop et al. (I972) Kelly & Tinsley (1973) Matthews (I969) Zweerink, Ito & Matsuhisa (I972) Kapuler et aL (197o) Kang & McAuslan (1972) Vilagines & McAuslan Ct971) Aubertin et al. (t97I) Burlingham et al. (1971) Kaplan, Wilbert & Black (I972)

Pox viruses

Herpes virus Paramyoxvirus Leukovirus Rhabdovirus Myxovirus Iridoviruses T2 virus Reoviruses Frog virus 3

Adenovirus Polyoma group (SV4o)

* References are not necessarily to the first report for a particular enzyme, contains only I. 3 ~ lipid (Bergold & Wellington, I954), although a more recent estimate gives 3"7 ~ (Bergold, I963).
The presence o f e n z y m e s in the virus particle

Members of the enveloped virus groups (except alpha and flavi) have been shown to contain enzymes concerned with nucleic acid metabolism (Table 5). Large viruses in five of the geometrical groups containing double-stranded nucleic acid (and one small virus of the polyoma group) have been shown to contain enzymes as part of their structure (Table 5)With one exception, none of the geometrical viruses containing single-stranded nucleic acid have been shown to contain enzymes. The exception is a parvovirus, Kilham rat virus (Salzman, I97I). However, there was no convincing demonstration that the D N A polymerase activity reported in the virus preparations was an integral part of the virus particle rather than a host contaminant.
W a t e r content

I f enveloped viruses bound, on average, 2"5 times as much water as geometrical did this would account for the average difference in Fig. I and 2 between the two classes (tenfold on

A classification using virus a n d g e n o m e s i z e

I43

Table 6. Water content or solvation o f some cells and viruses Water content (ml/gm dry matter) Hydrodynamic radius (nm) Cells
Escherichia coli (with wall) 3'0 E. coli (without wall) 4"0 Dialister pneumosintes 3"o

Cell or virus

References Davis et al. (I973) Davis et al. (I973) Loewy & Siekevitz 0969)

RSV AMV REO T7 PM2 Caulimovirus RI7 Off BSV TYMV TSV B component M component T component

2'70. 9 4"3!I'2

Enveloped viruses 74'o J.D. Harvey (personal communication) 8o-o J.D. Harvey (personal communication)

Geometrical viruses with double-stranded nucleic acid 1"5 49"0 J.D. Harvey (personal communication) 1-18+ o.o6 33"3 Camerini-Otero et al. (I974) 1. I ___ 0.08 33"0 Camerini-Otero et al. (I974) 1.7 28'4 J.D. Harvey (personal communication) Geometrical viruses with single-stranded nucleic acid Camerini-Otero et al. (1974) I'O2_O'O9 I4-o Camerini-Otero et al. (I974) I'22+O'O8 15"I Camerini-Otero et al. (1974) O'75+_O'O4 I7'2 J. D. Harvey (personal communication) o'78 14'7 J. D. Harvey (personal communication) 1'o9 17-4 J. D. Harvey (personal communication) I'I2 I6-2 J. D. Harvey (personal communication) I'26 15"4

a dry mass basis and 25-fold on a particle volume basis). The 20 to 30 % (on dry weight) of lipid would make a contribution of only a few per cent to the hydrated volume of the enveloped viruses. The water content of viruses has not received a great deal of attention. However, with the development of new methods for the rapid and accurate measurement of hydrodynamic radii new informaton is becoming available (Table 6). The error in these values is large even for the recent measurements and many more viruses require study. Nevertheless, the enveloped particles that have been studied have a water content like that of prokaryotic cells. Most of the geometrical virus particles for which data are available have about half as much water per unit dry matter.
Multiple genomes within a single envelope

A proportion of particles containing more than one genome within a single membrane has been found for the following groups of enveloped viruses: pox, herpes, paramyxo, myxo, leuko and alpha viruses (Simon, I972). The phenomenon appears not yet to have been established for the rhabdo- and flaviviruses. In general the phenomenonen does not occur in the geometrical viruses, although the long rods of the F f bacteriophage provide an analogous phenomenon (Simon, I972 ) and some baculoviruses may contain up to about 2o nucleocapsids within one outer protein membrane.
Exceptions

On the size data at present available, the alpha and flavi groups occupy an intermediate position as indicated in Fig. I and 2. They are also anomalous in that the virus particles do not appear to contain enzymes concerned with nucleic acid metabolism.

144

R . E . F . MATTHEWS
A. Enveloped viruses Double-stranded Single-stranded

/\
Pox
viruses

DNA Herpes
viruses

RNA No DNA step in

replication cycle

--...
DNA step in replication cycle

-....
to mRNA viruses

genome = m R N A

genome complementary

/
Alphaviruses

~
Flavi-

viruses

Rhabdo- Paramyxoviruses viruses

Myxo- RNA tumour

viruses

B. Geometrical viruses Double-stranded nucleic acid particles) Single-stranded nucleic acid

DNA RNA

/\

lcosahedral particles

/\

Helical rods

/\

DNA RNA

DNA RNA

Fig. 3. Summary of suggested divisions for groups of viruses.

Some cultures of tobacco ringspot virus (TRSV) contain a satellite virus (S-TRSV) which uses the TRSV coat protein built into a shell identical to that of TRSV. S-TRSV belongs to the geometrical class since it does not have a lipoprotein envelope. However, it is anomalous with respect to its nucleic acid. The R N A of S-TRSV has a mol. wt. of 1"2 Io 5 and I2 to 25 pieces of this size are encapsulated within a TRSV protein shell (Schneider, Hull & Markham, 1972; Sogo, Schneider & Koller, I974). N o other example of this situation is known. S-TRSV may represent a satellite 'viroid' which uses the protein shell of the virus upon which it is dependent. The high resistance to u.v. inactivation (Diener, Schneider & Smith, I974) supports the view that the amount of R N A required for infectivity is very small.
DISCUSSION

On the basis of the data outlined above, I suggest that the primary division of the viruses should be into two classes: A. Enveloped virus particles with the following properties, which resemble those of prokaryotic cells: (I) Bounded by a lipoprotein membrane. (2) A particle volume per unit of genetic material of the order of 2 IO5 nma[I@ daltons of nucleic acid, and a dry particle mass of 4 x lO8 daltons, per lO6 daltons of nucleic acid. (3) A water content in the range 3 to 4 ml/g dry matter.

A classification using virus and genome size

I45

(4) Presence of enzymes concerned with nucleic acid metabolism, except in togaviruses. (5) Occasional occurrence of more than one genome within a single envelope.

B. Geometrical virus particles with the following properties: (I) No limiting lipoprotein membrane. (2) A particle volume per unit weight of nucleic acid about one twenty-fifth, and a dry particle mass per unit weight of nucleic acid about one-tenth, that of the enveloped class. (3) Following from (2) they will have an expected water content of about 1.5 ml/g dry matter. (4) The absence of enzymes from the virus particle except for the larger members of the class. (5) No multiple genomes within a single envelope (except for some baculoviruses). Further subdivisions of the two classes
A grouping of animal viruses according to the strategy of their genome has been proposed (Baltimore, i97i ; Subak-Sharpe, I97I) and has been recently reviewed for the RNA viruses by Shatkin (i974). The enveloped viruses containing single-stranded RNA are readily subdivided according to the strategy of their genome (Fig. 3A). As noted earlier, the geometrical viruses appear to fall naturally into three subclasses. These suggestions are summarized in Fig. 3 B. The differences between the enveloped and geometrical classes can be highlighted by considering influenza virus. Although it is about three orders of magnitude smaller than Eseherichia coli it has a similar dry mass and particle volume per unit of nucleic acid. On the other hand influenza virus contains about the same amount of RNA as enteroviruses and yet has roughly Io times the dry mass and 20 to 30 times the particle volume. A proportion of the increased dry mass will be due to the presence of the lipoprotein envelope, but substantial amounts of other proteins must be present as well. There appears to be no biological or structural reason why such additional proteins should be a necessary consequence of the presence of a bilayer membrane. The greater dry mass per unit of nucleic acid may therefore be regarded as an independent property. However, a larger proportion of water in enveloped viruses might well result directly from the presence of a membrane with osmotic properties. Several lines of investigation could further test the validity of the classification. These include: (i) Careful re-examination of the size in solution (using laser light beat spectroscopy) of various virus groups, such as herpes-, alpha- and flaviviruses. Substantially larger particles than the published estimates could be expected. (ii) Accurate determination of the water content of many virus particles for which information is lacking at present. (iii) Re-investigation of the DNA polymerase activity reported for Kilham rat virus. (iv) Further structural studies on the icosahedral cytoplasmic deoxyriboviruses and the baculoviruses to delineate more clearly to which class they belong. A primary division of viruses into the two classes outlined above would have greater predictive value than current schemes and might perhaps correspond more closely to evolutionary origins. Wildy (1973) drew attention to the long tradition of separatism of the four main' branches' of virology, and he gave a summary of current information on possible bridging groups of viruses. A classification based on the divisions indicated in Fig. 3 would give substantial unity to virology as a whole, as it relegates categories based on host organisms to a subsidiary position.
io VIR 27

146

R.E.F.

MATTHEWS J. L o n g w o r t h a n d J. A . R

I w i s h t o t h a n k A . R . B e l l a m y , J. D . H a r v e y , J. K a l m a k o f f , Miles for most useful comments and discussion.

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(Received 3o October ~974)