Investigating the viral suppressor HC-pro inhibiting small rna methylation through functional comparison of HEN1 in angiosperm and bryophyte
Journal
Viruses
Journal Volume
13
Journal Issue
9
Date Issued
2021
Author(s)
Sanobar N
Lin P.-C
Pan Z.-J
Fang R.-Y
Tjita V
Chen F.-F
Wang H.-C
Tsai H.-L
Wu S.-H
Shen T.-L
Chen Y.-H
Abstract
In plants, HEN1-facilitated methylation at 3′ end ribose is a critical step of small-RNA (sRNA) biogenesis. A mutant of well-studied Arabidopsis HEN1 (AtHEN1), hen1-1, showed a defective developmental phenotype, indicating the importance of sRNA methylation. Moreover, Marchantia polymorpha has been identified to have a HEN1 ortholog gene (MpHEN1); however, its function remained unfathomed. Our in vivo and in vitro data have shown MpHEN1 activity being comparable with AtHEN1, and their substrate specificity towards duplex microRNA (miRNA) remained consistent. Furthermore, the phylogenetic tree and multiple alignment highlighted the conserved molecular evolution of the HEN1 family in plants. The P1/HC-Pro of the turnip mosaic virus (TuMV) is a known RNA silencing suppressor and inhibits HEN1 methylation of sRNAs. Here, we report that the HC-Pro physically binds with AtHEN1 through FRNK motif, inhibiting HEN1’s methylation activity. Moreover, the in vitro EMSA data indicates GST-HC-Pro of TuMV lacks sRNA duplex-binding ability. Surprisingly, the HC-Pro also inhibits MpHEN1 activity in a dosage-dependent manner, suggesting the possibility of interaction between HC-Pro and MpHEN1 as well. Further investigations on understanding interaction mechanisms of HEN1 and various HC-Pros can advance the knowledge of viral suppressors. ? 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Subjects
HEN1
HEN1-HC-Pro interaction
Methylation
SRNA
Viral suppressors
Arabidopsis protein
microRNA
recombinant protein
small interfering RNA
cysteine proteinase
HC-Pro protein, potyvirus
HEN1 protein, Arabidopsis
methyltransferase
plant protein
plant RNA
protein binding
viral protein
amino acid sequence
angiosperm
Arabidopsis
Article
binding affinity
biogenesis
chemoluminescence
controlled study
DNA methylation
enzyme specificity
Escherichia coli
female
gel mobility shift assay
gene
gene duplication
gene silencing
immunization
in vitro study
in vivo study
male
molecular evolution
mosaicism
moss
MpHEN1 gene
nonhuman
nucleosome
phenotype
phylogenetic tree
phylogeny
plant growth
plant immunity
polyacrylamide gel electrophoresis
protein degradation
protein expression
protein motif
protein purification
RNA extraction
RNA methylation
sequence alignment
Western blotting
chemistry
genetics
Marchantia
metabolism
methylation
Potyvirus
protein domain
virology
Amino Acid Motifs
Arabidopsis Proteins
Cysteine Endopeptidases
Methyltransferases
MicroRNAs
Phylogeny
Plant Proteins
Protein Binding
Protein Domains
Recombinant Proteins
RNA, Plant
Substrate Specificity
Viral Proteins
Type
journal article