title: The 20S proteasome α5 subunit of Arabidopsis thaliana carries an RNase activity and interacts in planta with the lettuce mosaic potyvirus HcPro protein.
abstract: In plants, the ubiquitin/26S proteasome system (UPS) plays a central role in protein degradation and is involved in many steps of defence mechanisms, regardless of the types of pathogen targeted. In addition to its proteolytic activities, the UPS ribonuclease (RNase) activity, previously detected in 20S proteasome preparations from cauliflower and sunflower (Helianthus annuus), has been shown to specifically target plant viral RNAs in vitro. In this study, we show that recombinant Arabidopsis thaliana proteasomal α(5) subunit expressed in Escherichia coli harbours an RNase activity that degrades Tobacco mosaic virus (TMV, Tobamovirus)- and Lettuce mosaic virus (LMV, Potyvirus)-derived RNAs in vitro. The analysis of mutated forms of the α(5) subunit demonstrated that mutation of a glutamic acid at position 110 affects RNase activity. Furthermore, it was demonstrated, using a bimolecular fluorescence complement assay, that the multifunctional helper component proteinase (HcPro) of LMV, already known to interfere with the 20S proteasome RNase activity in vitro, can interact in vivo with the recombinant α(5) subunit. Further experiments demonstrated that, in LMV-infected lettuce cells, α(5) is partially relocalized to HcPro-containing infection-specific inclusions. Susceptibility analyses of Arabidopsis mutants, knocked out for each At-PAE gene encoding α(5) , showed that one (KO-pae1) of the two mutants exhibited a significantly increased susceptibility to LMV infection. Taken together, these results extend to A. thaliana α(5) the range of HcPro-interacting proteasomal subunits, and suggest that HcPro may modulate its associated RNase activity which may contribute to an antiviral response.
authors.affiliation: Interactions Plante-Virus, UMR GDPP 1090, INRA Université de Bordeaux 2, BP 81, F-33883 Villenave d'Ornon Cedex, France.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Dielen
authors.forename: Flavio Tetsuo
authors.initials: FT
authors.lastname: Sassaki
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Walter
authors.forename: Thierry
authors.initials: T
authors.lastname: Michon
authors.forename: Guillaume
authors.initials: G
authors.lastname: Ménard
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Pagny
authors.forename: Renate
authors.initials: R
authors.lastname: Krause-Sakate
authors.forename: Ivan De Godoy
authors.initials: Ide G
authors.lastname: Maia
authors.forename: Saloua
authors.initials: S
authors.lastname: Badaoui
authors.forename: Olivier
authors.initials: O
authors.lastname: Le Gall
authors.forename: Thierry
authors.initials: T
authors.lastname: Candresse
authors.forename: Sylvie
authors.initials: S
authors.lastname: German-Retana
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000821:virology; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli; D018506:Gene Expression Regulation, Plant; D055785:Gene Knockdown Techniques; D018698:Glutamic Acid / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018545:Lactuca; D009154:Mutation / Q000235:genetics; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D021122:Protein Subunits; D012367:RNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D018698:Glutamic Acid; C559598:PAE1 protein, Arabidopsis; C559599:PAE2 protein, Arabidopsis; D012260:Ribonucleases; D003546:Cysteine Endopeptidases; C072775:HC-Pro protein, potyvirus; D046988:Proteasome Endopeptidase Complex