Tobacco mosaic virus

M.M. Hernandez

Tobacco mosaic virus

TMV
Genus Tobamovirus
15 members
naked, rigid rod,
+ unsegmented ss RNA

No family assignation by ICTV

STRUCTURAL FEATURES OF TMV

CAPSID
30 nm

Helical symmetry

18 nm

2 nm

RNA genome

NC protein
Images:P. Sforza, 2001
www.ppws.vt.edu/~sforza/tmv/tmv.html

TMV genome organization

6,395 nt
Capsid CP

Replicase RdRp
5 cap

183K
126 K

30 K
UAG (leaky)

MT-Hel

17.6 K

Movement MP

tRNAhis

Host: Nicotiana tabaccum

(Solanaceous ) & other 9 plant families
susceptible too

Disease:Mosaic (calico)

Mosaic disease on Tobacco leaf

Chlorosis

Stunting
Necrosis

Mosaic disease: significant losses in yield & $ value

TMV Life cycle

5 cap

Host Rb

a) Virus entry trough

abrasions on plant
tissue. Inside cell
associates with ER
b) spontaneous release
of few capsid (CP)
subunits 5' end of
genome is uncovered
c) Host ribosome
attaches to viral RNA,
moves down displacing
more CP units /
Modification of a diagram from: L. Stannard, Department of Medical Microbiology, U. of Cape Town

TMV Life cycle

(cont.)

d) Ribosome meets start

codon, translates first two
proteins (126K ,183 K) while
uncoating continues
co-traslational disassembly
e) 126 K ( MET-Hel) & 183 K
( RdRp) use viral RNA as
template to make full length
complementary neg. strand
RNA
/

Modification of a diagram from: L. Stannard, Department of Medical Microbiology, U. of Cape Town

TMV Life cycle

RdRp

+ Strand (genome)

(cont.)

Neg. strand

f) Neg. RNA strand used

by viral replicase
(RdPp/MET-Hel ) as
template for +RNA
g) Also, neg. RNA strand
has internal promoters
used by replicase to
make mRNA for 30K
protein (MP) and 17.5 K
(CP)

promoters

Transcription by RdRp

MP mRNA

CP mRNA

TMV Life cycle

(cont.)

h) MP combines
with viral +RNA
to move it into
new plant cells
through
plasmodesmata

http:www.sunysb.edu/biochem/BIOCHEM/facultypages/citovsky/TMV-TransportModel-III2.jpg

TMV Life cycle

(cont.)

i) Accumulation of +RNA
& CP proteins stimulates
assembly of progeny
virions

Viral RNA interacts with

discs

CP units self assemble

into discs

Electrostatic interaction stabilizes structure

into helix

M. Olvera de la Cruz

TMV Life cycle

(cont.)

j) massive TMR replication

occur in the X-bodies
(viroplasmas)

Szecsi et al. 1999

Hamacher et al. 2003

TMV Life cycle

(cont.)

k) Virus will remain in plant tissue and sap until delivered to

a new plant host usually by mechanical means

TMV diagnosis
Symptoms
E.M. & Serology
RT-PCR & RFLP

Letschert et al. 2002

HTTP;//BAKERLAB.BERKELEY.EDU/?IMG=1

Current Research on TMV

Improving diagnostic
techniques
Characterization of
MP and viral
movement through
plant
Characterization of
the Helicase & RdRp

Questions?

REFERENCES
Agrios G. N. 1988. Plant Pathology Third Edition. Academic Press, Inc. San Diego
pp: 658663
Ashby J., E. Boutant, M. Seemanpillai, A. Sambade, C. Ritzenthaler, M. Heinlein. 2006. Tobacco
Mosaic Virus Movement Protein Functions as a structural Microtubule-Associated Protein. Journal of
Virology. 80(17):8329-8344
Bos L. 1999. Plant Viruses Unique and intriguing pathogens. Backhuys Publishers Leiden
Dovas C., K. Efthimious, N. I. Katis. 2004. Generic detection and differentiation of tobamoviruses by
a spot nested RT-PCR-RFLP using dI-contining primers along with homologous dG-containing primers.
Journal of Virological Methods. 117:137-144
Goelet P., Lomonossoff G. P., Blutler P. JG, Akam M. E., Gait M. J., and Karn J., 1982. Nucleotide
sequence of the tobacco mosaic virus RNA. Proc. Natl. Acad. Sci. U.S.A. 79: 5818-5822
Goregaoker S. P. and J. N. Culver. 2003. Oligomerization and Activity of the Helicase Domain of the
Tobacco Mosaic Virus 126- and 183-Kilodalton Replicase Proteins. Journal of Virology. 77 (6): 35493556
Hirashima K. and Y. Watanabe. 2001. Tobamovirus Replicase Coding Region is Involved in Cell-toCell Movement. Journal of Virology. 75 (18) 8831-8836
Hirashima K. and Y. Watanabe. 2003. RNA Helicase Doamin of Tobamovirus Replicase Executes
Cell-to-Cell Movement Possibly through Collaboration with its Nonconserved Region. Journal of
Virology. 77 (22) 12357- 12362
Horst K.R.(reviser), 2001 Westcott=s Plant Disease Handbook Six Edition. Kluwer Academic
Plublishers.Boston pp:511-513
Jacobi V., G.D. Bachand, R.C. Hamelin and J.D. Castello. 1998. Development of a multiplex
immunocapture RT-PCR assay for detection and differentiation of tomato and tobacco mosaic
tobamoviruses. Journal of Virological Methods 74 (2):167-178
International Committee on Taxonomy of Viruses (ICTV) 2002. ICTVdB Virus Descriptions.
71.0.1.0.001 Tobacco mosaic virus (vulgare strain; ssp. NC82 strain). Retrieved November 22, 2002,
from: http://www.ncbi.nih.gov/ICTVdb/ICTVdB/71010001.htm
Kawakami F. M., R.W. Kim, R.N. Beachy. 2006 Domains of tobacco mosaic virus movement protein
essential for its membrane association. Journal of General Virology 87(9):2699-7707

Khan J. A. and J. Dijkstra editors, 2002. Plant Viruses as Molecular Pathogens Food Product Press 7
An Imprint of the Haworth Press Inc. New York
Lazarowitz S. G., 2001. Plant Viruses In: Fundamental Virology Fourth Edition. Knipe D. and P.M.
Howley Editors. Lippincott Williams & Wilkins, Philadelphia. pp:377-429
Lesemann D.-E., S. Winters. 1998. Detection and specific identification of tobamoviruses in horticultural
crops by immunoelectron microscopical methods (IEM) and ELISA. Mitteilungender Biologischen
Bundesanstalt fur Land-und Forstwirtschaft 357
Letschert B., A Gunter, L. Dietrich-Eckhardt, P. Willingmann, C. Heinze., 2002. Detection and
differentiation of serologically cross-reacting tobamoviruses of economical importance by RT-PCR and
RT-PCR-RFLP. Journal of Virological Methods 106:1-10
Lucas G. B. , C.L. Campbell, and L.T. Lucas, 1985. Introduction to Plant Diseases Identification and
Management. The AVI Publishing Company, Inc. Westport Connecticut. pp 254-261
Martin J.H. , W.H. Leonard and D. L. Stamp 1976 Principles of Field Crop Production Third Edition
MacMills Publishing Co., INC. London
Matthews R.E.F., 1992. Fundamentals of Plant Virology Academic Press Inc San Diego
Matthews R.E.F., 1999 Plant Virology Third Edition. Academic Press Inc San Diego. pp:207-225
Osman T. A. M., and K.W. Buck. 2003 Identification of a Region of the Tobacco Mosaic Virus 126- and
183-Kilodalton Replication Proteins Which bind Specifically to the Viral 3'-Terminal tRNA-like Structure.
Journal of Virology 77(16 ) 8669-8675
Padmanadban M.S., Goregaoker S.P., S. Golem, H. Shiferaw, and J. N. Culver. 2005. Interaction of
the Tobacco Mosaic Virus Replicase Protein with the Aux/IAA Protein PAP1/IAA26 Is associated with
Disease Development. Journal of Virology. 79 (4) 2549 - 2558
Sacristan S., J. M. Malpica, A. Fraile, F. Garcia-Arenal. 2003. Estimation of Population Bottlenecks
during Systemic Movement of Tobacco Mosaic Virus in Tobacco Plants. Journal of Virology 77(18) 99069911
Seemanpillai M., R. Elamawi, C. Ritzenthhaler, M. Heinlein. 2006. Challenging the Role of
Microtubules in Tobacco Mosaic Virus Movement by Drug Treatment is Disputable. Journal of Virology
80(13): 6712-6715
Torok V.A. and J.W. Randles. 2001. Tobacco mosaic virus RNA as an internal control for duplex RTPCR assay of pea germaplsm. Australian Plant Pathology 30: 227-230
Zaitlin M., 2001. Tobamovirus in: The Spriger Index of Viruses. Tidona C. A. and G. Darai Editors.
Springer-Verlag, Berlin

1.SolutionofTMVCapsidProteins
Electrostaticrepulsioninhibits
+protein+proteinaggregation.

2. Add ssTMV+ RNA (assembly

infective virus): Electrostatic
attraction stabilizes viral assembly.

Salt(M)
ZRNA= -1/mer

3. Sphere-like (icosahedral) RNA virus

(Tsuruta et al)

Acidity
4. Capsid + Hole (density RNA crystal)
self-assembles from capsid protein + viral
RNA solution