Recent discoveries of the roles of RNA-based silencing during virus-host interactions of multicellular eukaryotes have revealed fundamental mechanisms that govern virus susceptibility and defense. Silencing mechanisms operate to condition local and systemic antiviral states in response to infection, but also to modulate cellular or viral functions that enable virus replication, invasiveness and latency. Understanding these mechanisms will illuminate basic events that occur at the interface between virus and host, and will reveal natural antiviral defense mechanisms that may be exploited for directed therapies or preventative measures. The genetic, genomic and technical aspects of the Arabidopsis model have proven exceptionally useful in revealing RNA silencing functions that limit virus infection at the cell-autonomous and cell-nonautonomous levels, as well as silencing functions that govern stress responses, development, and repressive chromatin. During the current project period, Arabidopsis was used to identify the roles of cellular and viral factors, such as virus-encoded suppressors of RNA silencing, during antiviral defense and counter defensive processes. This research, and work from several other groups, led to conceptualization of a three-phase model for antiviral silencing in Arabidopsis. This model describes events occurring during the initial targeting phase, the siRNA amplification phase and the systemic silencing phase. The three Aims of the proposed project focus on virus-host interactions that occur during each phase. First, new high-throughput sequencing technology will be used to analyze the genetic requirements for initial targeting of viral genomes by DICER-LIKE (DCL) factors during early stages of infection, and to test the hypothesis that initial targeting yields primarily (+)-sense siRNA that seed the subsequent amplification step. Second, the genetic requirements and detailed accumulation patterns of siRNA formed during the RNA- DEPENDENT RNA POLYMERASE6-dependent amplification phase will be determined, and the small RNA populations that interact with a viral RNA silencing suppressor will be identified. And third, the hypothesis that viral silencing suppressors function in vascular cells to inhibit cell-nonautonomous, DCL4-dependent systemic signals that limit virus invasiveness will be tested.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AI043288-16
Application #
8220939
Study Section
Virology - B Study Section (VIRB)
Program Officer
Park, Eun-Chung
Project Start
1997-09-30
Project End
2013-02-28
Budget Start
2012-03-01
Budget End
2013-02-28
Support Year
16
Fiscal Year
2012
Total Cost
$321,630
Indirect Cost
$125,610
Name
Donald Danforth Plant Science Center
Department
Type
DUNS #
044193006
City
St. Louis
State
MO
Country
United States
Zip Code
63132
Campo, Sonia; Gilbert, Kerrigan B; Carrington, James C (2016) Small RNA-Based Antiviral Defense in the Phytopathogenic Fungus Colletotrichum higginsianum. PLoS Pathog 12:e1005640
Carbonell, Alberto; Carrington, James C; Daròs, José-Antonio (2016) Fast-forward generation of effective artificial small RNAs for enhanced antiviral defense in plants. RNA Dis 3:
Fahlgren, Noah; Hill, Steven T; Carrington, James C et al. (2016) P-SAMS: a web site for plant artificial microRNA and synthetic trans-acting small interfering RNA design. Bioinformatics 32:157-8
Wang, Haifeng; Beyene, Getu; Zhai, Jixian et al. (2015) CG gene body DNA methylation changes and evolution of duplicated genes in cassava. Proc Natl Acad Sci U S A 112:13729-34
Fahlgren, Noah; Feldman, Maximilian; Gehan, Malia A et al. (2015) A Versatile Phenotyping System and Analytics Platform Reveals Diverse Temporal Responses to Water Availability in Setaria. Mol Plant 8:1520-35
Carbonell, Alberto; Fahlgren, Noah; Mitchell, Skyler et al. (2015) Highly specific gene silencing in a monocot species by artificial microRNAs derived from chimeric miRNA precursors. Plant J 82:1061-75
Carbonell, Alberto; Carrington, James C (2015) Antiviral roles of plant ARGONAUTES. Curr Opin Plant Biol 27:111-7
Garcia-Ruiz, Hernan; Carbonell, Alberto; Hoyer, J Steen et al. (2015) Roles and programming of Arabidopsis ARGONAUTE proteins during Turnip mosaic virus infection. PLoS Pathog 11:e1004755
Minoia, Sofia; Carbonell, Alberto; Di Serio, Francesco et al. (2014) Specific argonautes selectively bind small RNAs derived from potato spindle tuber viroid and attenuate viroid accumulation in vivo. J Virol 88:11933-45
Carbonell, Alberto; Takeda, Atsushi; Fahlgren, Noah et al. (2014) New generation of artificial MicroRNA and synthetic trans-acting small interfering RNA vectors for efficient gene silencing in Arabidopsis. Plant Physiol 165:15-29

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