7182 Nucleic Acids Research, Vol. 18, No. 23
AMV-capsid protein-gene
Nucleotide sequence of
H.Steinkellner, G.Himmler, D.Mattanovich and H.Katinger Institute of
Applied Microbiology, University
of
Peter JordanstraBe 82, 1190 Vienna,
Agriculture,
Austria
EMBL accession no. X55460
Submitted October 11, 1990 To prove that the coat protein gene of Arabis Mosaic Nepovirus (AMV) (C.M.I./A.A.B. Description of Plant Viruses No 16) is located at the 3 -end of RNA2, as it was shown for other Nepoviruses (1, 2), we sequenced the 3' end of the cDNA (3) of RNA2 of a grapevine isolate of AMy. Since the functional coat protein of nepoviruses is released from a polyprotein by posttranslational cleavage, the exact NH2-terminal end of the coat protein gene could only be determined by comparing the experimentally achieved NH2 amino acids of the coat protein (GLAGRGSVQVPKDCQAGIYL) with the deduced amino acid sequence of the cDNA. The corresponding sequence of the cDNA would give a hypothetical translational product of 505 amino acids (Mol. wt. 5509 Dalton), which is in good agreement with the Mol. wt. estimated by SDS-PAGE electrophoresis (4). The cleavage site is RIG, the same as it has been shown for grapevine fanleaf virus (1). Comparison with the coat protein of grapevine fanleaf virus and AMV shows a similarity of 70% (Alignment by Meyers and Miller's method) (5).
ACKNOWLEDGEMENTS This work is
excellent
CTT
60
50
TCA AGA TGCCA
ACC
A
70
so
90
GGC GGG AAT ATA TCT GAA AAC CCT TIGA TCT TAG AGA CAT GGT TAG A
G
I
Y
L
K
T
D
L
L
R
D
G
N
V
S
1.
Serghini,M.A., Fuchs,M., Pink,M., Reinbolt,J., (1990) Grapevine Fanleaf Virus: Sequence Analysis Location. J.
2.
Nucleotide mosaic 3.
4.
Sequence
nepovirus
Steinkellner,H.,
110
GaIl,O.
genetic organisation
and
of
Dunez,J.
(1989)
Hungarian grapevine
chrome
F
S
G
I
Q
Y
K
E
N
I
T
A
G
1
G
N
N
K
2
L
A
A
D
W
E
F
V
and MiIJer,W. (1988)
Biosci.
4, 11
Optimal alignment
150
140
m
P
N
F
170
160
K
V
V
I
R
Y
P
Igo
A
N
A
1150
1140
1130
T
T
S
T
V
A
820
840
830
L
Y
A
E
F
E
N
r
I
Y
A
A
K
S
F
C
L
K
L
190
c
y
I
E
E
D
G
S
F
A
I
E
I
R
T
G
210
200
I
T
W
V
M
S
F
220
D
A
Y
D
A
F
N
G
L
S
K
R
860
p
y
870
L
H
1:
aso
$90
G
240
230
250
270
AAT TAC TAG TAG TAT TTC AAC TAC TGC TAG TCC CGC GTA TAC
T
P
L
R
L
I
D
A
T
S
S
I
S
T
T
A
S
P
A
Y
T
ACT L
P
P
L
E
Y
D
L
S
A
T
S
590
s
S
K
280
290
3 10
V
S
L
L
L
G
0
T
V
L
s
S
F
930
920
T
9 so
S
T
L
N
F
Y
v
P
H
W
L
L
H
.
x
N
G
T
T
S
960
970
A
P
S
G
E
1000
CGG TAC TCA TAA GGT GTA TAA rIr TAA TAA TAC ACT CTT GAG 77A G
T
H
W
D
L
340
330
350
360
E
I
S
G
980
D
L
D
Y
G
E
L
C
G
H
A
M
W
F
K
V
Y
N
F
N
N
T
L
L
S
710
700
690
680
T
A
K
N
P
V
V
1020
1010
L
G
L
1260
E
G
F
G
X
s
S
1290
G
G
V
1300
I
X
T
L
C
I
C
G
I
v
K
G
K
V
E
720
H
V
C
C
D
K
A
T
G
m
D
G
1350
1340
1330
1320
V
S
TCG
TGT
TTG
TIGC
Trr
ACA
AAA
R
V
C
A
L
Q
N
E
I
A
L
P
D
L
S
1040
1050
D
Y
F
L
V
1060
W
V
D
P
S
S
C
E
T
E
1390
GCT ATA CAT AGG CAA rrA TGC GGG TGC TAA CCC AAA TAC TGC ATT
CTT
CCC
F
P
Y
I
G
N
y
A
1410
G
S
F
A
N
p
N
T
A
1430
1420
L
1440
1080
1070
F
G
1380
1370
ATG GCT TGC TAT TAA GCT
CAA TGA rrA 7rr CCT TT. GGT TGA CTT TTC ATC ATT CAC TGT -A N
M
Q
1030
AAT TGA AGA AAT TGC ACT TCC GGA TCr rrC TGT C-CC ATC
Y
CTA Trr AGG CAT TGG GGG TAT TGT CAA GGG AAA GGT GCA TGT TTG Y
K
990
S e
L
P
ATC CTT CTA TAG TCG ATG TGA CCT TGA CTA TGG AGA ACT CTG TGG TCA TGC TAT GTG GTT
D
1250
1280
1400 120
L
940
A
L s
I
CAT AGC CCC AAG TGG AGA AAC AGC GAA GAT GCC AGT ACT CGT GCA
D
670
660
650
640
GTC TGT CCC TCA TTG GC'r TCT GCA TCA TAA AAA TGG GAC CAC CTC
G
1240
1360 3 00
H
A
AGC ATC
Y
m
AAG CTC Trr TAC GAG TAC GTT GAA Trr TTA CGC AAT CTC AGG GCC
630
620
610
600
TGC CTA CAA GAG TGT GTC CCT GTT JkTT GGG ACA GAC TCT TGT TGA A
H
900
Q
A I
H
L
1230
1310 260
A
1210
1200
1190
TGC CCA TAT
TAC CAT CAC GAA ATT GTG TIGG TGA TAA GGC CAC TGG CAT GGA TGI;
I
T
F
NAT GTG GGA TTT GGA GGG TGA GTT TGG AAA GAG rrC TGG TGG TGT R
H
910
GTT TCT GAC CAT CCC ACC ACT GGA ATA TGA CTT AAG TGC AAC CAG r
TTT
CCC CTA CCA TCG AAC TCC ACT TCG Crr AAT TGA CGC GCA ATC GGC
580
570
S60
550
N
P
S
T
F
N
850
1270 CTT CAC AGG TAT TAC GTG GGT TAT GAG TTT TGA TGC TTA CAA CCG
D
GTG TTA CAT CGA GGA GGA TGG GAG Crr TGC TAT TGA AAT ACG CTC
540
530
520
S
C
GAT AGC TTG TCA TGG ACr CCA TCA TGG AAT CCT TGA TCT TAA GTT
510
Soo
L
111C
1160
TAC TAC TAG CAC TGT AGC CCT TIGG AGA CAA TCC
490
490
470
GCC CAA CTT CAT TTA TAG CTT GGA CGC Trr TAA TGG GTC TCT CAA p
Comp.
C10
C601 C9
m v
in linear space.
17.
1220 TGT AAT GCC AAA TTT TAA GGT GGT GAT TCG CTA TCC GGC CAA TGC
and
145, 191- 194.
Meyers,E.W.
V
CGA ATT ATA TGC TGA GTT TIGA GGC AGC AAA AAG 7`TT TCT TGG CAA
E
and
17, 7809-7819.
RNA2. Nuci. Acids Res.
A
G
PinlkCL.
and
Himm-ler,G., Laimer,M., Mattanovich,D., Bisztray,G.
4 50
440
430
42 0
410
460
TGG Grr rrC AGG TAT TCA GTA CGA AAA GM GAT CAC TGC TGG AAT
Walter,B&
and Coat Protein Cistron
GGCTC TCCTT TA GGCCC TAA GTT ACATTCAAGCT
130
120
for
1433-1441.
1180 100
Ruiker
Katinger,H. (1989) Konstruktion von cDNA von Arabis Mosaik Virus und deren Anwendung fiir Diagnose. Mitt. Klostemneuburg Nr. 6, 242 -246. Takemnoto,Y., Nagahara,Y., Fukuyama,K., Tsukihara,T. and Iwak,M. (1985) Crystallisation and Preliminary Characterization of Arabis Mosaic Virus. Virol.
5.
Virol. 71,
Gen.
Brault,V., Hibrand,L., Candresse,T., Le
CAC TGG AAA TAA CAA AGA ATr GGC GGC GGA TTIG GGA Grr CGT TGT T
and Florian
help.
C3 C TGAGGGrC
TG
Regina Sagl
We thank
REFERENCES
Applic.
GC/GG CT
BMLF grant No 480 and FFWF grant
supported by
No 7715-Bio.
T
V
D
L
Y
S
R
W
L
A
I
K
L
TG
D
TAA AGC CA-A
K
A
K
AMV-capsid protein-gene
Nucleotide sequence of
H.Steinkellner, G.Himmler, D.Mattanovich and H.Katinger Institute of
Applied Microbiology, University
of
Peter JordanstraBe 82, 1190 Vienna,
Agriculture,
Austria
EMBL accession no. X55460
Submitted October 11, 1990 To prove that the coat protein gene of Arabis Mosaic Nepovirus (AMV) (C.M.I./A.A.B. Description of Plant Viruses No 16) is located at the 3 -end of RNA2, as it was shown for other Nepoviruses (1, 2), we sequenced the 3' end of the cDNA (3) of RNA2 of a grapevine isolate of AMy. Since the functional coat protein of nepoviruses is released from a polyprotein by posttranslational cleavage, the exact NH2-terminal end of the coat protein gene could only be determined by comparing the experimentally achieved NH2 amino acids of the coat protein (GLAGRGSVQVPKDCQAGIYL) with the deduced amino acid sequence of the cDNA. The corresponding sequence of the cDNA would give a hypothetical translational product of 505 amino acids (Mol. wt. 5509 Dalton), which is in good agreement with the Mol. wt. estimated by SDS-PAGE electrophoresis (4). The cleavage site is RIG, the same as it has been shown for grapevine fanleaf virus (1). Comparison with the coat protein of grapevine fanleaf virus and AMV shows a similarity of 70% (Alignment by Meyers and Miller's method) (5).
ACKNOWLEDGEMENTS This work is
excellent
CTT
60
50
TCA AGA TGCCA
ACC
A
70
so
90
GGC GGG AAT ATA TCT GAA AAC CCT TIGA TCT TAG AGA CAT GGT TAG A
G
I
Y
L
K
T
D
L
L
R
D
G
N
V
S
1.
Serghini,M.A., Fuchs,M., Pink,M., Reinbolt,J., (1990) Grapevine Fanleaf Virus: Sequence Analysis Location. J.
2.
Nucleotide mosaic 3.
4.
Sequence
nepovirus
Steinkellner,H.,
110
GaIl,O.
genetic organisation
and
of
Dunez,J.
(1989)
Hungarian grapevine
chrome
F
S
G
I
Q
Y
K
E
N
I
T
A
G
1
G
N
N
K
2
L
A
A
D
W
E
F
V
and MiIJer,W. (1988)
Biosci.
4, 11
Optimal alignment
150
140
m
P
N
F
170
160
K
V
V
I
R
Y
P
Igo
A
N
A
1150
1140
1130
T
T
S
T
V
A
820
840
830
L
Y
A
E
F
E
N
r
I
Y
A
A
K
S
F
C
L
K
L
190
c
y
I
E
E
D
G
S
F
A
I
E
I
R
T
G
210
200
I
T
W
V
M
S
F
220
D
A
Y
D
A
F
N
G
L
S
K
R
860
p
y
870
L
H
1:
aso
$90
G
240
230
250
270
AAT TAC TAG TAG TAT TTC AAC TAC TGC TAG TCC CGC GTA TAC
T
P
L
R
L
I
D
A
T
S
S
I
S
T
T
A
S
P
A
Y
T
ACT L
P
P
L
E
Y
D
L
S
A
T
S
590
s
S
K
280
290
3 10
V
S
L
L
L
G
0
T
V
L
s
S
F
930
920
T
9 so
S
T
L
N
F
Y
v
P
H
W
L
L
H
.
x
N
G
T
T
S
960
970
A
P
S
G
E
1000
CGG TAC TCA TAA GGT GTA TAA rIr TAA TAA TAC ACT CTT GAG 77A G
T
H
W
D
L
340
330
350
360
E
I
S
G
980
D
L
D
Y
G
E
L
C
G
H
A
M
W
F
K
V
Y
N
F
N
N
T
L
L
S
710
700
690
680
T
A
K
N
P
V
V
1020
1010
L
G
L
1260
E
G
F
G
X
s
S
1290
G
G
V
1300
I
X
T
L
C
I
C
G
I
v
K
G
K
V
E
720
H
V
C
C
D
K
A
T
G
m
D
G
1350
1340
1330
1320
V
S
TCG
TGT
TTG
TIGC
Trr
ACA
AAA
R
V
C
A
L
Q
N
E
I
A
L
P
D
L
S
1040
1050
D
Y
F
L
V
1060
W
V
D
P
S
S
C
E
T
E
1390
GCT ATA CAT AGG CAA rrA TGC GGG TGC TAA CCC AAA TAC TGC ATT
CTT
CCC
F
P
Y
I
G
N
y
A
1410
G
S
F
A
N
p
N
T
A
1430
1420
L
1440
1080
1070
F
G
1380
1370
ATG GCT TGC TAT TAA GCT
CAA TGA rrA 7rr CCT TT. GGT TGA CTT TTC ATC ATT CAC TGT -A N
M
Q
1030
AAT TGA AGA AAT TGC ACT TCC GGA TCr rrC TGT C-CC ATC
Y
CTA Trr AGG CAT TGG GGG TAT TGT CAA GGG AAA GGT GCA TGT TTG Y
K
990
S e
L
P
ATC CTT CTA TAG TCG ATG TGA CCT TGA CTA TGG AGA ACT CTG TGG TCA TGC TAT GTG GTT
D
1250
1280
1400 120
L
940
A
L s
I
CAT AGC CCC AAG TGG AGA AAC AGC GAA GAT GCC AGT ACT CGT GCA
D
670
660
650
640
GTC TGT CCC TCA TTG GC'r TCT GCA TCA TAA AAA TGG GAC CAC CTC
G
1240
1360 3 00
H
A
AGC ATC
Y
m
AAG CTC Trr TAC GAG TAC GTT GAA Trr TTA CGC AAT CTC AGG GCC
630
620
610
600
TGC CTA CAA GAG TGT GTC CCT GTT JkTT GGG ACA GAC TCT TGT TGA A
H
900
Q
A I
H
L
1230
1310 260
A
1210
1200
1190
TGC CCA TAT
TAC CAT CAC GAA ATT GTG TIGG TGA TAA GGC CAC TGG CAT GGA TGI;
I
T
F
NAT GTG GGA TTT GGA GGG TGA GTT TGG AAA GAG rrC TGG TGG TGT R
H
910
GTT TCT GAC CAT CCC ACC ACT GGA ATA TGA CTT AAG TGC AAC CAG r
TTT
CCC CTA CCA TCG AAC TCC ACT TCG Crr AAT TGA CGC GCA ATC GGC
580
570
S60
550
N
P
S
T
F
N
850
1270 CTT CAC AGG TAT TAC GTG GGT TAT GAG TTT TGA TGC TTA CAA CCG
D
GTG TTA CAT CGA GGA GGA TGG GAG Crr TGC TAT TGA AAT ACG CTC
540
530
520
S
C
GAT AGC TTG TCA TGG ACr CCA TCA TGG AAT CCT TGA TCT TAA GTT
510
Soo
L
111C
1160
TAC TAC TAG CAC TGT AGC CCT TIGG AGA CAA TCC
490
490
470
GCC CAA CTT CAT TTA TAG CTT GGA CGC Trr TAA TGG GTC TCT CAA p
Comp.
C10
C601 C9
m v
in linear space.
17.
1220 TGT AAT GCC AAA TTT TAA GGT GGT GAT TCG CTA TCC GGC CAA TGC
and
145, 191- 194.
Meyers,E.W.
V
CGA ATT ATA TGC TGA GTT TIGA GGC AGC AAA AAG 7`TT TCT TGG CAA
E
and
17, 7809-7819.
RNA2. Nuci. Acids Res.
A
G
PinlkCL.
and
Himm-ler,G., Laimer,M., Mattanovich,D., Bisztray,G.
4 50
440
430
42 0
410
460
TGG Grr rrC AGG TAT TCA GTA CGA AAA GM GAT CAC TGC TGG AAT
Walter,B&
and Coat Protein Cistron
GGCTC TCCTT TA GGCCC TAA GTT ACATTCAAGCT
130
120
for
1433-1441.
1180 100
Ruiker
Katinger,H. (1989) Konstruktion von cDNA von Arabis Mosaik Virus und deren Anwendung fiir Diagnose. Mitt. Klostemneuburg Nr. 6, 242 -246. Takemnoto,Y., Nagahara,Y., Fukuyama,K., Tsukihara,T. and Iwak,M. (1985) Crystallisation and Preliminary Characterization of Arabis Mosaic Virus. Virol.
5.
Virol. 71,
Gen.
Brault,V., Hibrand,L., Candresse,T., Le
CAC TGG AAA TAA CAA AGA ATr GGC GGC GGA TTIG GGA Grr CGT TGT T
and Florian
help.
C3 C TGAGGGrC
TG
Regina Sagl
We thank
REFERENCES
Applic.
GC/GG CT
BMLF grant No 480 and FFWF grant
supported by
No 7715-Bio.
T
V
D
L
Y
S
R
W
L
A
I
K
L
TG
D
TAA AGC CA-A
K
A
K
