Académique Documents
Professionnel Documents
Culture Documents
JOURNAL OF VIROLOGY
June 2006, p. 5841–5853
GEMINIVIRUS
• PLANT PATHOGEN
• LEGUMINOUS PLANTS
• AGRO-ECONOMIC LOSS
A A T
A
T T
A T Replication initiation /
A Rep nicking site
T
C C
G C
A T
C G
T A
C G
A T
C G
G C
G C
G C CGCGACCGGTGTATTG
ATCGGTGTACAC//TCGGTGTATCGGTGTCTTA//GCCCATAG
Replication Interference Assay
I
Criterion for motif classification:
I
I
1. Motifs include atleast 5 members
N,I 2. Members interact with CaAL1
I 3. Aas typically involved in Pro-Pro
I interactions
4. Motifs are related to plant protein
N
Significance of Aptamer Approach for Developing
Virus Resistant Plants
1. Stringent in vivo screening most likely selects peptides that fold
correctly, stably expressed and bind with high affinity
3. Small size enables the peptides to move passively into the nucleus
?
1. How does virus get the replication machinary in a differentiated cell?
2. What factors induce the host cell to synthesize the proteins required for
virus?
3. What factors direct the host replication machinary to the work on viral
genome?
Thus, proper understanding of geminiviral replication
enables us to control its pathogenesis at the replication
step and also lets us understand the plant replication
mechanism
AC3 IS THE REPLICATION ENHANCER
Unbound Bound
(400mM Salt Wash) Unbound Bound
(600mM Salt Wash)
1 L A+ P B M
e nol a P B M
PB M
PB M
AL3 interaction with Rep: in vitro binding assay
MBP+AL1
MBP+AL1
MBP+AL1
AL3+AL1
AL3+AL1
AL3+AL1
AL1
AL1
AL1
Bound Fractions
AL3 Oligomerization: Sucrose Gradient Ultracentrifugation
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
AC2
Rep/AC1 REn/AC3
Effect of AC3 interacting Host proteins on
Replication of Viral genome: Yeast Model
• TBY-2 protoplast transfection with wild type and mutant AC3 constructs
CR AC1
TBY-2