Potyvirus Helper Component-Proteinase Self-Interaction in the Yeast Two-Hybrid System and Delineation of the Interaction Domain Involved

Urcuqui-Inchima, Silvio; Walter, Jocelyne; Drugeon, Gabrièle; German-Retana, Sylvie; Haenni, Anne‐Lise; Candresse, Thierry; Bernardi, Françoise; Gall, Olivier Le

doi:10.1006/viro.1999.9725

Cited by 51 publications

(50 citation statements)

References 31 publications

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“…Moreover, HCPro and P1b are alike in their ability to self-interact ( Fig. 4 and 5) (23,54,62,70,74), to bind siRNAs ( Fig. 6 and 7) (37), and to enhance PVX pathogenicity ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Protease Activity, Self Interaction, and Small Interfering RNA Binding of the Silencing Suppressor P1b fromCucumber Vein Yellowing Ipomovirus

Vallí

Dujovny

Garcı́a

2008

J Virol

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The RNA silencing pathway mediated by small interfering RNAs (siRNAs) plays an important antiviral role in eukaryotes. To counteract this defense barrier, a large number of plant viruses express proteins with RNA silencing suppression activity. Recently, it was reported that the ipomovirus Cucumber vein yellowing virus (CVYV), which lacks the typical silencing suppressor of members of the family Potyviridae, i.e., HCPro, has a duplicated P1 coding sequence and that the downstream P1 copy, named P1b, has silencing suppression activity. In this study, we provide experimental evidence that P1b is a serine protease that self-cleaves at its C terminus but that its proteolytic activity is not essential for silencing suppression. In contrast, a putative zinc finger and a conserved basic motif in the N-terminal region of the protein are required for efficient silencing suppression. In vitro gel filtration-fast protein liquid chromatography and in vivo bimolecular fluorescence complementation assays showed that P1b binds itself to form oligomeric structures and that the zinc finger-like motif is essential for the self interaction. Moreover, we observed that CVYV P1b forms complexes with synthetic siRNAs, and this ability correlated with both silencing suppression activity and enhancement of Potato virus X pathogenicity in a mutational analysis. Together, these results suggest that CVYV P1b resembles potyviral HCPro and other viral proteins in interfering RNA silencing by preventing siRNA loading into the RNAinduced silencing complex.

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“…Moreover, HCPro and P1b are alike in their ability to self-interact ( Fig. 4 and 5) (23,54,62,70,74), to bind siRNAs ( Fig. 6 and 7) (37), and to enhance PVX pathogenicity ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Protease Activity, Self Interaction, and Small Interfering RNA Binding of the Silencing Suppressor P1b fromCucumber Vein Yellowing Ipomovirus

Vallí

Dujovny

Garcı́a

2008

J Virol

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“…As a negative control of interaction, we used the murine laminin ␥1 sequence (Chang et al, 1996) cloned into pLexA in combination with the different recombinant pGAD3S2X. The quantitative ␤-galactosidase assays were performed according to Urcuqui-Inchima et al (1999).…”

Section: Two-hybrid Detection Of Protein-protein Interactionsmentioning

confidence: 99%

A New C-Type Cyclin-Dependent Kinase from Tomato Expressed in Dividing Tissues Does Not Interact with Mitotic and G1 Cyclins

et al. 2001

Self Cite

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Cyclin-dependent kinases (CDKs) form a conserved superfamily of eukaryotic serine-threonine protein kinases whose activity requires the binding of a cyclin protein. CDKs are involved in many aspects of cell biology and notably in the regulation of the cell cycle. Three cDNAs encoding a C-type CDK, and a member of each B-type CDK subfamily, were isolated from tomato (Lycopsersicon esculentum Mill.) and designated Lyces;CDKC;1 (accession no. AJ294903), Lyces; CDKB1;1 (accession no. AJ297916), and Lyces;CDKB2;1 (accession no. AJ297917). The predicted amino acid sequences displayed the characteristic PITAIRE (CDKC), PPTALRE (CDKB1), and PPTTLRE (CDKB2) motives in the cyclin-binding domain, clearly identifying the type of CDK. The accumulation of all transcripts was associated preferentially with dividing tissues in developing tomato fruit and vegetative organs. In contrast to that of CDKA and CDKBs, the transcription pattern of Lyces;CDKC;1 was shown to be independent of hormone and sugar supply in tomato cell suspension cultures and excised roots. This observation, together with the absence of a patchy expression profile in in situ hybridization experiments, suggests a non-cell cycle regulation of Lyces;CDKC;1. Using a two-hybrid assay, we showed that Lyces;CDKC;1 did not interact with mitotic and G1 cyclins. The role of plant CDKCs in the regulation of cell division and differentiation is discussed with regard to the known function of their animal counterparts.

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“…HC-Pro, a silencing suppressor protein, is also known as a synergy factor needed for potyvirus infection [1,18,31,32]. HC-Pro is a multifunctional protein, which helps in transmission of the virus through aphids, replication and systemic movement of the virus [13,39]. p25 of PVX is known as a silencing suppressor protein.…”

Section: Introductionmentioning

confidence: 99%

Expression of symptoms, viral coat protein and silencing suppressor gene during mixed infection of a N–Wi strain of potato virus Y and an asymptomatic strain of potato virus X

Senanayake

Mandal

2014

VirusDis.

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Potato virus Y (PVY) and potato virus X (PVX), the RNA viruses of two different genera results into synergistic interactions on mixed infection. In this study, a N-Wi strain of PVY and a PVX strain that is asymptomatic on potato were used to study their interactions during mixed infection in Nicotiana benthamiana and Nicotiana tabacum with reference to symptom expression, level of coat protein (CP) using ELISA and suppressor gene using real time PCR under high temperature (26-40°C) and low temperature (5-25°C) conditions. Both mixed and single infection caused severe necrosis and death of N. benthamiana plants. Single infection of these viruses in N. tabacum showed mild symptoms but mixed infection caused more severe symptoms. Synergistic symptoms were more pronounced under low temperature conditions than at high temperature. In low temperature conditions, the CP level of PVX in N. benthamiana was twofold higher than PVY and both the viruses reached at peak at 28 dpi in single virus infection. When PVY and PVX inoculated together, the CP levels of both the viruses increased and reached to the peak earlier (within 7-14 days) than that in the single virus inoculation. Although, the CP level of PVX was higher than PVY in mixed infection, at later stage (28 dpi) both the CP level declined to the similar level. The level of p25 suppressor gene was higher than HCPro in single inoculation. However, under mixed inoculation of PVY and PVX, expression of p25 was declined to the level of HC-Pro when the CP levels of both the virus also were observed to decline. The expression pattern of CP and suppressor gene was different in plants when mixed infection was created by inoculation of one virus followed by the other. This study showed the level of CP and suppressor gene of specific strain of PVY and PVX during their mixed infection in tobacco.

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Potyvirus Helper Component-Proteinase Self-Interaction in the Yeast Two-Hybrid System and Delineation of the Interaction Domain Involved

Cited by 51 publications

References 31 publications

Protease Activity, Self Interaction, and Small Interfering RNA Binding of the Silencing Suppressor P1b fromCucumber Vein Yellowing Ipomovirus

Protease Activity, Self Interaction, and Small Interfering RNA Binding of the Silencing Suppressor P1b fromCucumber Vein Yellowing Ipomovirus

A New C-Type Cyclin-Dependent Kinase from Tomato Expressed in Dividing Tissues Does Not Interact with Mitotic and G1 Cyclins

Expression of symptoms, viral coat protein and silencing suppressor gene during mixed infection of a N–Wi strain of potato virus Y and an asymptomatic strain of potato virus X

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