Kayayyaki

Sunan furotin na ORF45 na Kaposi's sarcoma-associated herpesvirus (KSHV) shine takamaiman furotin tegument na musamman-gammaherpesvirus.Nazarin mu da suka gabata sun bayyana mahimmin matsayinsa a farkon farkon da ƙarshen matakan kamuwa da KSHV.A cikin wannan binciken, mun bincika hulɗar ORF45 ta amfani da panel na ƙwayoyin rigakafi na monoclonal.Baya ga abubuwan da aka gano a baya an gano kinase mai kayyadewa (ERK) da sunadaran p90 ribosomal S6 kinase (RSK), mun sami wasu sunadaran da aka goge da yawa, gami da fitattun abubuwan ~ 38 kDa da ~ 130 kDa.Mass spectrometry ya bayyana cewa sunadaran 38-kDa shine kwayar cutar hoto ta ORF33 kuma furotin 130-kDa shine salon salula USP7 (protease na musamman na kowane wuri 7).Mun tsara yankin dauri na ORF33 zuwa amino acid na carboxyl-terminal 19 (aa) na ORF45 da yankin dauri na USP7 zuwa dalilin yarda da aka ruwaito a yankin tsakiyar ORF45.Yin amfani da tabo na immunofluorescence, mun lura da colocalization na ORF45 tare da ORF33 ko USP7 duka a ƙarƙashin ions yanayi masu canzawa kuma a cikin kwayoyin cutar KSHV.Haka kuma, mun lura da komawar ORF45-dogara na wani yanki na ORF33/USP7 daga tsakiya zuwa cytoplasm.Mun gano cewa ORF45 ya haifar da karuwa a tarin furotin na ORF33 wanda aka soke idan ko dai ORF33- ko USP7-daure yanki a cikin ORF45 an share.Bugu da ƙari kuma, shafewar da aka adana na carboxyl ta ORF45 a cikin KSHV genome ya rage girman matakin furotin na ORF33 a cikin ƙwayoyin cuta na KSHV da kuma kawar da samar da ƙwayoyin cuta.Gabaɗaya, sakamakonmu ba wai kawai ya bayyana sabbin abubuwan haɗin gwiwar ORF45 ba, har ma ya nuna cewa hulɗar tsakanin waɗannan sunadaran suna da mahimmanci ga maimaitawar KSHV.KSHV ORF45 furotin ne mai aiki da yawa wanda ake buƙata don maimaita KSHV lytic, amma ainihin hanyoyin da ORF45 ke aiwatar da ayyukansa masu mahimmanci ba su da tabbas.Nazarinmu na baya sun bayyana cewa duk ORF45 pr otein a cikin sel suna wanzuwa a cikin ma'aunin nauyi mai nauyi.Don haka mun nemi siffata ma'amala ta ORF45 don ba da haske game da ayyukanta yayin kwafin lytic.Yin amfani da rukunin ƙwayoyin rigakafi na monoclonal, mun bincika hulɗar ORF45 a cikin sel masu kamuwa da KSHV.Mun gano sabbin abokan haɗin gwiwa guda biyu na ORF45: furotin ORF33 mai hoto ko bidiyo mai zagaya yanar gizo da sauri da takamaiman protease 7 (USP7).Mun ƙara nuna cewa hulɗar tsakanin ORF45 da ORF33 yana da mahimmanci don samar da ingantaccen ƙwayoyin ƙwayoyin cuta na KSHV, yana nuna cewa tsangwama da aka yi niyya tare da wannan hulɗar na iya wakiltar wani sabon dabara don hana KSHV lytic kwafi.