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1.
J Virol ; 81(23): 12872-80, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17898061

ABSTRACT

Viroids are small, circular, single-stranded RNA molecules that, while not coding for any protein, cause several plant diseases. Viroids rely for their infectious cycle on host proteins, most of which are likely to be involved in endogenous RNA-mediated phenomena. Therefore, characterization of host factors interacting with the viroid may contribute to the elucidation of RNA-related pathways of the hosts. Potato spindle tuber viroid (PSTVd) infects several members of the Solanaceae family. In an RNA ligand screening we have previously isolated the tomato protein Virp1 by its ability to specifically interact with PSTVd positive-strand RNA. Virp1 is a bromodomain-containing protein with an atypical RNA binding domain and a nuclear localization signal. Here we investigate the role of Virp1 in the viroid infection cycle by the use of transgenic lines of Nicotiana tabacum and Nicotiana benthamiana that either overexpress the tomato Virp1 RNA or suppress the orthologous Nicotiana genes through RNA silencing. Plants of the Virp1-suppressed lines were not infected by PSTVd or Citrus exocortis viroid through mechanical inoculation, indicating a major role of Virp1 in viroid infection. On the other hand, overexpression of tomato Virp1 in N. tabacum and N. benthamiana plants did not affect PSTVd KF 440-2 infectivity or symptomatology in these species. Transfection experiments with isolated protoplasts revealed that Virp1-suppressed cells were unable to sustain viroid replication, suggesting that resistance to viroid infection in Virp1-suppressed plants is likely the result of cell-autonomous events.


Subject(s)
Nicotiana/virology , Plant Proteins/physiology , RNA-Binding Proteins/physiology , Viroids/physiology , DNA, Plant/chemistry , DNA, Plant/genetics , Gene Expression , Gene Silencing , Molecular Sequence Data , Plant Diseases/virology , Plant Proteins/genetics , Plants, Genetically Modified/virology , RNA-Binding Proteins/genetics , Sequence Analysis, DNA , Nicotiana/genetics
2.
Plant J ; 12(3): 615-23, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9351246

ABSTRACT

The Arabidopsis thaliana MALE STERILITY 2 (MS2) gene product is involved in male gametogenesis. The first abnormalities in pollen development of ms2 mutants are seen at the stage in microsporogenesis when microspores are released from tetrads. Expression of the MS2 gene is observed in tapetum of wild-type flowers at, and shortly after, the release of microspores from tetrads. The MS2 promoter controls GUS expression at a comparable stage in the tapetum of transgenic tobacco containing an MS2 promoter-GUS fusion. The occasional pollen grains produced by mutant ms2 plants have very thin pollen walls. They are also sensitive to acetolysis treatment, which is a test for the presence of an exine layer. The MS2 gene product shows sequence similarity to a jojoba protein that converts wax fatty acids to fatty alcohols. A possible function of the MS2 protein as a fatty acyl reductase in the formation of pollen wall substances is discussed.


Subject(s)
Arabidopsis Proteins , Arabidopsis/enzymology , Plant Proteins/genetics , Amino Acid Sequence , Arabidopsis/genetics , Arabidopsis/physiology , Base Sequence , Molecular Sequence Data , Phenotype , Plant Proteins/drug effects , Plant Proteins/metabolism , Promoter Regions, Genetic
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