M1 RNA is important for the in-cell solubility of its cognate C5 protein: Implications for RNA-mediated protein folding.

It is one of the fundamental questions in biology how proteins efficiently fold into their native conformations despite off-pathway events such as misfolding and aggregation in living cells. Although molecular chaperones have been known to assist the de novo folding of certain types of proteins, the role of a binding partner (or a ligand) in the folding and in-cell solubility of its interacting protein still remains poorly defined. RNase P is responsible for the maturation of tRNAs as adaptor molecules of amino acids in ribosomal protein synthesis. The RNase P from Escherichia coli, composed of M1 RNA and C5 protein, is a prototypical ribozyme in which the RNA subunit contains the catalytic activity. Using E. coli RNase P, we demonstrate that M1 RNA plays a pivotal role in the in-cell solubility of C5 protein both in vitro and in vivo. Mutations in either the C5 protein or M1 RNA that affect their interactions significantly abolished the folding of C5 protein. Moreover, we find that M1 RNA provides quality insurance of interacting C5 protein, either by promoting the degradation of C5 mutants in the presence of functional proteolytic machinery, or by abolishing their solubility if the machinery is non-functional. Our results describe a crucial role of M1 RNA in the folding, in-cell solubility, and, consequently, the proteostasis of the client C5 protein, giving new insight into the biological role of RNAs as chaperones and mediators that ensure the quality of interacting proteins.

The C5 protein of euphorbia leaf curl virus is a virulence factor and gene silencing suppressor.

The genome of a monopartite begomovirus, or the DNA-A component of a bipartite begomovirus, typically encodes six proteins: two on the viral strand (AV1/V1 and AV2/V2) and four on the complementary strand (AC1/C1, AC2/C2, AC3/C3, AC4/C4). Recent studies, however, have identified additional begomoviral proteins with various functions. This paper reports that euphorbia leaf curl virus (EuLCV), a monopartite begomovirus, encodes a seventh protein, C5. Promoter activity of the upstream fragment of the EuLCV C5 gene was shown using a GUS expression vector. EuLCV C5 also enhanced the pathogenicity and accumulation of potato virus X (PVX) in Nicotiana benthamiana. Localization studies revealed that EuLCV C5 localizes to the cytoplasm and nucleus, forming granular structures on the cell membrane. Additionally, C5 acts as a post-transcriptional gene silencing (PTGS) suppressor. A C5 deletion mutant of EuLCV (EuLCV-ΔC5) exhibited reduced pathogenicity and viral accumulation compared to wild-type EuLCV in N. benthamiana.

The C5 protein encoded by Ageratum leaf curl Sichuan virus is a virulence factor and contributes to the virus infection.

Earlier reports have indicated that begomoviruses encode four proteins (AC1/C1, AC2/C2, AC3/C3, and AC4/C4 proteins) using complementary-sense DNA as the template. In recent years, several reports have shown that some begomoviruses also encode an AC5/C5 protein from the complementary DNA strand, and these AC5/C5 proteins play different roles in virus infections. Here, we provide evidence showing that Ageratum leaf curl Sichuan virus (ALCScV), a monopartite begomovirus, also encodes a C5 protein that is important for disease symptom formation and can affect viral replication. Infection of Nicotiana benthamiana plants with a potato virus X (PVX)-based vector carrying the ALCScV C5 gene resulted in more severe disease symptoms and higher virus accumulation levels. ALCScV C5 protein can be found in the cytoplasm and the nucleus. Furthermore, this protein is also a suppressor of posttranscriptional gene silencing. Mutational analysis showed that knockout of C5 gene expression significantly reduced ALCScV-induced disease symptoms and virus accumulation, while expression of the C5 gene using the PVX-based vector enhanced ALCScV accumulation in coinfected N. benthamiana plants.