Table 1 Potential functionally relevant amino acid substitutions in HeV proteins

P/V/W

R149C

Within STAT1-binding region; could modulate IFN antagonism.

LD / A

STAT1-binding assays⁸⁶

P/V/W

G219E

Within amyloidogenic motif; could influence fibrillization, proposed to be linked to immune evasion.

LD / C, D

Amyloid formation studies³³

P/V/W

D304G

Within amyloidogenic motif; could influence fibrillization and immune evasion.

LD / A

Amyloid formation studies³³

P/V/W

P352S

Alters local flexibility; flanks a conserved helical segment predicted to be a protein-binding motif.

LD / B

Structural modelling (Supplementary Discussion)

P/V/W

H139Y

Within STAT1-binding region; could modulate IFN antagonism.

non-LD / D

STAT1-binding assays⁸⁶

P/V/W

R170G

Near exportin-1-binding region; could influence subcellular localization.

non-LD / A to D

Exportin-1 binding assays and mutagenesis³²

F

A141T

Lies within the heptad repeat A; could influence fusion efficiency.

non-LD / D

Mutagenesis assays⁸⁷

G

S175T

Located within a predicted T-cell epitope; may alter antigenicity or immune evasion.

LD / D

NiV immunogenic epitope mapping studies³²

G

R248G

Near K246, which plays a critical role in fusion triggering; could affect syncytium formation and fusion efficiency.

non-LD / C

NiV G-F-binding screening and mutagenesis⁸⁸

G

N306K

N-glycosylation site; probably affects fusion activity

non-LD / B

Mutagenesis assays⁸⁹

L

Q321H

Adjacent to putative nucleotide NTP entry channel; could influence NTP affinity.

LD / C, D

Comparative structural analysis with cryo-EM structures of the NiV RNA polymerase^90,91

L

H2081R

Near conserved regions involved in RNA capping; could indirectly influence RNA capping performance.

LD / A

Comparative structural analysis with cryo-EM structures of the NiV L^90,91

L

I2110T

Near conserved regions involved in RNA capping; could indirectly influence RNA capping performance.

LD / A

Comparative structural analysis with cryo-EM structures of the NiV L^90,91