3.4.22.45 A387E no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 C16A substitution mutants introduced into amino-proximal region of helper component protease HC-Pro, vector transmission abolished 683105 3.4.22.45 C16AC20A double mutant generated, vector transmission abolished 683105 3.4.22.45 C20A no effects on vector transmission 683105 3.4.22.45 C20R no effects on vector transmission, randomized substitution 683105 3.4.22.45 C291S inactive in aphid transmission 137295 3.4.22.45 C305S active in aphid transmission, similar to wild-type 137295 3.4.22.45 C344A/H417A the mutant shows abolished self-cleavage 732424 3.4.22.45 C390W no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 C46A vector transmission abolished 683105 3.4.22.45 C49A vector transmission abolished 683105 3.4.22.45 C49AC46A double mutant generated, vector transmission abolished 683105 3.4.22.45 C649S active site mutant, no activity 137288 3.4.22.45 C694S mutant is proteolytically active and capable of cell-to-cell and long-distance movements 137288 3.4.22.45 D193Y mutants of helper component protease HC-Pro of clover yellow vein virus, generated by site-directed mutagenesis or by PCR-based primer extension mutagenesis, revertant of D193Y also created, single mutant alone does not induce lethal necrosis 683106 3.4.22.45 D421N no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 D446A mutation does not abolish in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 D458G no effect on zucchini yellow mosaic virus symptoms 711038 3.4.22.45 D506Y very mild symptoms of zucchini yellow mosaic virus infection 711038 3.4.22.45 D715E mutant is proteolytically active and capable of cell-to-cell and long-distance movements 137288 3.4.22.45 E273A no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 E299A/D300A RNA silencing suppressor activity reduced compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 E360A/D361A RNA silencing suppressor activity reduced compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows mild etching 698120 3.4.22.45 E396N affects symptom severity and viral pathogenicity 713072 3.4.22.45 E443K RNA silencing suppressor activity increased compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 E450A mutation does not abolish in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 E452D RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 F205L affects symptom severity and viral pathogenicity 713072 3.4.22.45 H429A mutation does not abolish in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 H722S active site mutant, no activity 137288 3.4.22.45 I11L RNA silencing suppressor activity increased compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 I225V defective in aphid transmission activity 137294 3.4.22.45 I33 mutants of helper component protease HC-Pro of clover yellow vein virus generated by site-directed mutagenesis or by PCR-based primer extension mutagenesis 683106 3.4.22.45 I442M no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 K307E active in aphid transmission 137295 3.4.22.45 K307H inactive in aphid transmission 137295 3.4.22.45 K307Q inactive in aphid transmission 137295 3.4.22.45 K307R active in aphid transmission 137295 3.4.22.45 K309N RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 K432A mutation does not abolish in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 K452A mutation does not abolish in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 K454T RNA silencing suppressor activity increased compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 K50E defective in aphid transmission activity 137294 3.4.22.45 K51E defective in aphid transmission activity 137294 3.4.22.45 K52E/K52A the mutations disrupt interaction between the enzyme and aphids 732424 3.4.22.45 K7N no effects on vector transmission, randomized substitution 683105 3.4.22.45 L134H the mutant shows abolished RNA-silencing suppression 732424 3.4.22.45 additional information a point mutation in the highly conserved FRNK box produces the HC-ProFINK protein that shows no sRNA binding 718356 3.4.22.45 additional information certain mutations introduced to the short, variable region of the enzyme increase accumulation of the enzyme and prolong the time which it remains effective in suppression of RNA silencing 732407 3.4.22.45 additional information construction of deletion mutants of the enzyme with 5'-proximal deletions of 12 to 720 nucleotides, mutant infection of wheat analysis, overview 670997 3.4.22.45 additional information deletion of the first 93 residues of the N-terminus of ZYMV HC-Pro. Mutation of the conserved Gly456 residue does not affect the autoproteolytic activity of ZYMV HC-Pro 717316 3.4.22.45 additional information enzyme replacements, overview 670090 3.4.22.45 additional information generation of different N- and C-terminal truncated DELTA HC-Pro constructs, i.e. comprising residues 93-456, 114-456, and 139-456, or 1-322, 1-350, and 1-372, construction of recombinant mutant MBP:HA-HC-ProFRNK, MBP:HAHC-ProFINK, and truncated MBP:HA-HC-ProFRNK. HC-ProFRNK/FINK clearly inhibits Arabidopsis thaliana HEN1 activity in vitro. HC-ProFRNK/FINK, but not the truncated proteins HC-Pro 93-456, and HC-Pro 1-372 displaying decreased in vitro affinity for AtHEN1 binding, inhibits the methyltransferase activity of AtHEN1 in vitro 717924 3.4.22.45 additional information inactivation by point mutation HCL134H, inhibits synergism and RNA silencing activity 670522 3.4.22.45 additional information mutant generation within coding region of helper component protease HC-Pro protein, nine synonymous substitutions and 15 of 25 non-synonymous substitutions shown to be without phenotypic effect, four non-synonymous substitutions, including one reverting to wild type, shown to disply attenuated systemic infection, six non-synonymous substitutions and one nonsense substitution shown to display abolished systemic infectivity 683717 3.4.22.45 additional information mutants used in compensatory evolution experiments: E299A (0.6127 RNA-silencing suppression activity relative to wild type, numeration corresponds to the location in the full polyprotein), E360A/D361A (0.1507 RNA-silencing suppression activity relative to wild type, numeration corresponds to the location in the full polyprotein), V192A (0.7388 RNA-silencing suppression activity relative to wild type, numeration corresponds to the location in the full polyprotein), Y423H (1.1020 RNA-silencing suppression activity relative to wild type, numeration corresponds to the location in the full polyprotein), E443K (1.2631 RNA-silencing suppression activity relative to wild type, numeration corresponds to the location in the full polyprotein), K454T (1.1592 RNA-silencing suppression activity relative to wild type, numeration corresponds to the location in the full polyprotein), mutations fixed during compensatory evolution experiments are also characterized by RNA-silencing suppression activity values 712985 3.4.22.45 additional information mutations in the 4E binding site of HCpro reducing virulence of PVA -, 718061 3.4.22.45 additional information mutations of a short variable region affect interactions with a microtubule-associated protein and induce necrotic responses in tobacco leaves -, 732542 3.4.22.45 additional information random-insertion scanning mutagenesis, generation of an pentapeptide-insertion scanning mutagenesis library of helper component protease HC-Pro, performed by using the mutation generation system F701 MGS, pentapeptide scanning mutants in pBIN61S-HC-Pro mapped by restriction digestion, insertion position determined by sequencing -, 683718 3.4.22.45 additional information three deletion mutants designed for HC-Pro protein of potato virus Y, HC-Pro1 (residues 98 to 456), HC-Pro2 (residues 1 to 298) and HC-Pro3 (residues 1 to 97) used for yeast two-hybrid analysis -, 683832 3.4.22.45 N193Y RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 N194D RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 N19I no effects on vector transmission, randomized substitution 683105 3.4.22.45 N200S RNA silencing suppressor activity increased compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 P310A/T311A the mutations disrupt interaction between the enzyme and capsid protein 732424 3.4.22.45 P311L RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 Q265H RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 R127G no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 R165G RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 R180I affects symptom severity and viral pathogenicity 713072 3.4.22.45 R180I/E396N induces transient leaf mottling, affects symptom severity and viral pathogenicity 713072 3.4.22.45 R180I/F205L/E396N the mutant is completely defective in its capacity to block miRNA regulation 713072 3.4.22.45 R240A/K241A/H242A no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 R247A/K248A no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 R455A mutation abolishes in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 R45K no effects on vector transmission, randomized substitution 683105 3.4.22.45 R51I mutants of helper component protease HC-Pro of clover yellow vein virus generated by site-directed mutagenesis or by PCR-based primer extension mutagenesis 683106 3.4.22.45 RK182,4AA the mutant shows abolished RNA-silencing suppression 732424 3.4.22.45 RK246,7AA the mutant shows abolished RNA-silencing suppression 732424 3.4.22.45 RRH234,5,6AAA the mutant shows abolished self-cleavage 732424 3.4.22.45 S610T mutant is proteolytically active and capable of cell-to-cell and long-distance movements 137288 3.4.22.45 T27I mutants of helper component protease HC-Pro of clover yellow vein virus generated by site-directed mutagenesis or by PCR-based primer extension mutagenesis, revertant of T27I also created, single mutant alone does not induce lethal necrosis but retains ability to induce necrotic symptoms 683106 3.4.22.45 T27I/D193Y double mutant 683106 3.4.22.45 T310A mutation abolishes in vitro interaction between helper-component-proteinase and coat protein 713074 3.4.22.45 V135A no RNA silencing suppressor activity, Nicotiana benthamiana plant inoculated with infectious transcript shows no etching 698120 3.4.22.45 V192A RNA silencing suppressor activity reduced compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows mild etching 698120 3.4.22.45 V355L RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 V419A RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 Y234H RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120 3.4.22.45 Y309A defective in aphid transmission activity 137294 3.4.22.45 Y344S RNA silencing suppressor activity similar to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows mild etching 698120 3.4.22.45 Y423H RNA silencing suppressor activity increased compared to wild-type, Nicotiana benthamiana plant inoculated with infectious transcript shows etching 698120