# Molecular Population Genetics of Aspen Mosaic-Associated Virus in Finland and Sweden
## 3 Results and Discussion
### Complete Sequence of AsMaV-RNA1, RNA2 and RNA 5 of Distant Isolates
The N-terminal region of the GPP precursor of AsMaV contains a signal peptide cleavage site, located at amino acid position 22 and 23 (AYT\(\downarrow\)SIR) (SignalP-6.0 Prediction [[https://services.healthtech.dtu.dk/services/SignalP-6.0/](https://services.healthtech.dtu.dk/services/SignalP-6.0/)]([https://services.healthtech.dtu.dk/services/SignalP-6.0/](https://services.healthtech.dtu.dk/services/SignalP-6.0/)), 12 April 2023). The similar cleavage site has been reported for PPSMV-1 at the same position but different sequence (AMA\(\downarrow\)S) [63]. The glycoprotein contains four transmembrane helices (TMH) at amino acid positions 2-24, 106-128, 170-187, and 580-599 (TMHMM v.2.0 on [[https://services.healthtech.dtu.dk/services/TMHMM-2.0/](https://services.healthtech.dtu.dk/services/TMHMM-2.0/)]([https://services.healthtech.dtu.dk/services/TMHMM-2.0/](https://services.healthtech.dtu.dk/services/TMHMM-2.0/)), 12 April 2023). However, six glycosylation sites have been reported for emaraviruses, AsMaV had five out of six sites at the amino acid position 62, 201, 239, 328, and 438 which were predicted by NetNGlyc-1.0 ([[https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/](https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/)]([https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/](https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/)), 12 April 2023). Interestingly, the Mucin-type O-glycosylation site which is a widespread post-translational modification of proteins found in the entire animal kingdom, but also in higher plants, has been detected in divergent isolates of AsMaV identified in this study (E58627 and E58632) at the amino acid position 549. This was not detected in the previously published isolate of AsMaV. Interestingly, Mucin-type O-glycosylation sites have been detected in actinidia virus-2 (AcV-2), another species of clade A emaraviruses, in the positions 209, 254, and 264. It should be mentioned that Mucin-type O-glycosylation is an evolutionarily conserved protein modification reported for membrane-bound and -secreted proteins which are involved in protein secretion, stability, processing, and function [64]. More analyzing should be performed to understand the effects of these variations. The AsMaV RNA5 was 1324 nts in size in sequenced isolates of E58627 and E58632. In both isolates, the ORF5 initiated with a start codon of AUG at the positions 70-72 and terminated with a stop codon of UGA at the positions 811-813 encoding the P28 (247 aa) with unknown function. There was only one aa change in the divergent isolates in comparison to the reference isolate (E55809), S\({}_{240}\) instead of N\({}_{240}\). These sequences shared 98% and 99-100% identity with E55089 and each other, respectively, at nt and amino acid levels. It has been reported that the P28 of AsMaV shares a similar structure with ABC proteins [6]. Figure 4: Motifs reported for RdRP of emaraviruses from phylogenetic clade A. SDR–1, SDR–2, and SDR–3 are the three main regions reported for RdRP in emaraviruses. SDR–1 domain contains the endonuclease motifs and might be involved in “cap–snatching”. SDR–2 contains the polymerase motifs designated motifs A–E, and SDR–3 is a Cap-binding domain.