Title Analysis of ADAR protein during productive HSV-1 Infection Title (english) Analyza ADAR proteina tijekom produktivne infekcije HSV-1 Author Justina Oluchi Echeta Mentor Igor Jurak (mentor)Committee member Igor Jurak (predsjednik povjerenstva)Committee member Jurica Novak (član povjerenstva)Committee member Nicholas Bradshaw (član povjerenstva)Granter University of Rijeka Defense date and country 2022-07-27, Croatia Scientific / art field, BIOTECHNICAL SCIENCES Abstract ADAR proteins are enzymes involved in the editing of dsRNA transcripts by
deamination in which adenosine is changed to Inosine. This alteration
ultimately affects protein translation. ADAR1 has two isoforms; ADAR1p110
and ADAR1p150, ADAR2 and ADAR3 make up the mammalian ADAR family.
Both isoforms of ADAR1 and ADAR2 are catalytically active while ADAR3 is
not. ADARs edit both self and non-self dsRNA. Several studies have shown
the role of ADAR proteins mostly in RNA viruses, however, the role of these
proteins in dsDNA viruses is largely unknown. During viral infection, they can
play an antiviral role by forming part of the cell’s innate immune response, a
proviral role by blocking viral dsRNA from being recognized by the immune
sensors, thereby facilitating viral replication in the host cell, or both roles as
observed in Influenza A virus.
Based on in vitro experiments, we have established cell models to analyse
the role of ADAR proteins during productive HSV-1 infection. ADAR protein
and mRNA levels during productive HSV-1 infection were determined,
showing upregulation of ADAR during the infection. We observed an
upregulation of ADAR proteins in U2OS, HeLa, HEK293T, and HFF cells
infected with wild-type HSV-1 using MOIs 5 and 10, but not with MOI of 1.
Interestingly, we observed ADAR cells-specific response to infection with UV
inactivated HSV-1, which we cannot explain at this point. To investigate the
functional relevance of ADAR1 and ADAR2 proteins for virus replication, we
transfected cells with ADAR1 and ADAR2 expressing plasmids to overexpress
ADAR in cells. Surprisingly, we did not observe any effect on virus DNA
replication, indicating that ADAR1 and ADAR2 might not play a role in
productive HSV-1 infection. However further investigation is needed to
answer this question.
Abstract (croatian) ADAR protein su enzimi uklučeni u uređivanje dsRNA transkripata
deaminacijom u kojoj se adenozin mijenja u inozin. Ova promjena u
konačnici može utjecati na stabilnost RNA te na translaciju proteina. Obitelj
ADAR proteina čine ADAR1, koji ima dvije izoforme: ADAR1p110 i
ADAR1p150, ADAR2 i ADAR3. ADAR1 i ADAR2 su katalitički aktivni dok
ADAR3 nije. ADAR uređuju vlastite i brojne druge dsRNA. Nekoliko je studija
pokazalo ulogu proteina ADAR većinom u RNA virusima, , međutim njihova
uloga kod infekcije s dsDNA virusima slabo je istražena. Tijekom virusne
infekcije, oni mogu imati antivirusnu ulogu čineći dio urođenog imunološkog
odgovora stanica, ali i provirusnu ulogu, blokirajući virusnu dsRNA da je
prepozna imunološki senzor, čime se olakšava replikacija virusa u stanici
domaćinu, ili obje uloge kao što je primijećeno kod Influenza A virus.
Na temelju in vitro eksperimenata, uspostavili smo stanične modele za
analizu uloge ADAR proteina tijekom productivene HSV-1 infekcije. Određene
su razine ADAR proteina i mRNA tijekom produktivne HSV-1 infekcije. Uočili
smo pojačanu regulaciju ADAR u U2OS, HeLa, HEK293T i HFF stanicama
zaraženim divljim tipom HSV-1 te je takva pojačana ekspresija ovisila o
količini virusa kojom se inficira stanica. Zanimljivo je da smo uočili različiti
ADAR odgovor na infekciju s UV inaktiviranim virusom kod različitih vrsta
stanica, što u ovom trenutku ne možemo objasniti. Nadalje, kako bismo
istražili funkcionalnu relevantnost ADAR1 i ADAR2 proteina za replikaciju
virusa, transfecirali smo stanice s ADAR1 i ADAR2 ekspresijskim plazmidima
za prekomjernu ekspresiju ADAR-a. Naime, nismo primijetili nikakav učinak
na replikaciju DNA virusa, što ukazuje da ADAR1 i ADAR2 možda nemaju
ulogu u produktivnoj HSV-1 infekciji. Međutim, potrebna su daljnja
istraživanja kako bi se odgovorilo na ova pitanja.
Keywords
ADAR
HSV-1
dsRNA
infection
innate immune response
Keywords (english)
ADAR
HSV-1
dsRNA
infekcija
urođeni imunološki odgovor
Language english URN:NBN urn:nbn:hr:193:745757 Study programme Title: Biotechnology for the Life Sciences Type of resource Text File origin Born digital Access conditions Open access Terms of use Created on 2022-10-24 12:27:44
