Nā huahana

E like me nā maʻi maʻi RNA maikaʻi ʻē aʻe, ua hana hou ʻia nā tombusvirus e ka membrane-bound viral replicase complex (VRC).Aia ka VRC i ka p92 virus-coded RNA-dependent RNA polymerase (RdRp), ka viral p33 RNA chaperone, a me kekahi mau protein host i koho ʻia.I mea e lilo ai i RdRp hana ma hope o kona unuhi ʻana, pono e hoʻokomo ʻia ka protein replication p92 i loko o ka VRC, a ma hope o kona hoʻāla ʻana.Ua hōʻike mua mākou i loko o kahi hōʻike hoʻoheheʻe hū ʻole i ka cell-free yeast extract-based assay e pono ai ka hoʻōla ʻana o ka Tomato bushy stunt virus (TBSV) RdRp i kahi mea hoʻokipa hiki ke hoʻoheheʻe ʻia.Ma keia 'atikala, 'ike mākou i ka cellular heat shock protein 70 (Hsp70) ma ke 'ano he mea ho'okipa i koho 'ia no ka ho'ola 'ana i ka TBSV RdRp recombinant N-terminally truncated recombinant.Eia kekahi, ʻo ka hoʻopaʻa ʻana i ka hana Hsp70 liʻiliʻi i hoʻokumu ʻia i ka RNA synthesis e ka tombusvirus RdRp in vitro.Eia kekahi, hōʻike mākou i nā phospholipid neutral, ʻo ia hoʻi, phosphatidylethanolamine (PE) a me phosphatidylcholine (PC), hoʻonui i ka hoʻoulu ʻana o RdRp in vitro.Ma ka hoʻohālikelike ʻana, hōʻike ka phosphatidylglycerol (PG) i kahi hopena paʻa ikaika a mana i ka hoʻoulu ʻana o RdRp in vitro.Hōʻike pū mākou i ka hoʻoulu ʻana o PE a me PC i ka pilina RdRp-viral plus-strand RNA [(+)RNA], ʻoiai ʻo PG ke kāohi i ka hoʻopaʻa ʻana o ka RNA viral i ka RdRp.Ma muli o ka hoʻoulu ʻana i nā hana hoʻoweliweli o nā phospholipids like ʻole i ka tombusvirus RdRp activation, ke manaʻo nei mākou e hoʻohana ʻia ka hui lipid o nā membrane subcellular i manaʻo ʻia e nā tombusviruses e hoʻoponopono i ka hui VRC hou i ka wā o ka maʻi. ʻO ka virus-coded RNA-dependent RNA polymerase (RdRp), nona ke kuleana no ka hoʻohui ʻana i ka progeny RNA viral i loko o nā keʻena maʻi o kekahi mau maʻi RNA strand maikaʻi, ʻaʻole i hana mua.Hiki paha i kēia hoʻolālā ke pale aku i ka synthesis RNA viral i loko o ka cytosol e alakaʻi koke i ka hoʻokomo ʻana i nā pane antiviral cellular RNA.I ka wā o ka hui ʻana o ka paʻa paʻa paʻa me ka membrane, ua hoʻāla ʻia ka RdRp viral ma o kahi kaʻina hana maopopo ʻole e hiki ai i ka RdRp ke hana i ka RNA synthesis.Ma ka hoʻohana ʻana iā TBSV RdRp, hōʻike mākou i ka hui pū ʻana o ka cellular Hsp70 chaperone a me nā phospholipid neutral e hoʻoikaika i ka hoʻoulu ʻana o RdRp in vitro.I ka hoʻokaʻawale, phosphatidylglycerol (PG) loaʻa i ka hopena inhibitory nui i ka in vitro RdRp activation a me RdRp-viral RNA pilina, e hōʻike ana i ka microdomain membranous e hoʻopuni ana i ka RdRp e hoʻopilikia nui i kona hiki no ka RNA synthesis.No laila, ʻo ka hoʻoulu ʻana o ka RdRp viral e hilinaʻi ʻia i nā mea hoʻokipa lehulehu i loko o nā maʻi maʻi.