Identification of interaction domains in the pseudorabies virus ribonucleotide reductase large and small subunits.
Alphaherpesvirus
RNR
dNTPs
pUL39
pUL40
Journal
Veterinary microbiology
ISSN: 1873-2542
Titre abrégé: Vet Microbiol
Pays: Netherlands
ID NLM: 7705469
Informations de publication
Date de publication:
Jul 2020
Jul 2020
Historique:
received:
23
02
2020
revised:
16
05
2020
accepted:
25
05
2020
entrez:
2
7
2020
pubmed:
2
7
2020
medline:
19
3
2021
Statut:
ppublish
Résumé
Alphaherpesviral ribonucleotide reductase (RNR) is composed of large (pUL39, RR1) and small (pUL40, RR2) subunits. This enzyme can catalyze conversion of ribonucleotide to deoxynucleotide diphosphates that are further phosphorylated into deoxynucleotide triphosphate (dNTPs). The dNTPs are substrates for de novo viral DNA synthesis in infected host cells. The enzymatic activity of RNR depends on association between RR1 and RR2. However, the molecular basis underlying alphaherpesviral RNR complex formation is still largely unknown. In the current study, we investigated the pseudorabies virus (PRV) RNR interaction domains in pUL39 and pUL40. The interaction of pUL39 and pUL40 was identified by co-immunoprecipitation (co-IP) and colocalization analyses. Furthermore, the interaction amino acid (aa) domains in pUL39 and pUL40 were mapped using a series of truncated proteins. Consequently, the 90-210 aa in pUL39 was identified to be responsible for the interaction with pUL40. In turn, the 66-152, 218-258 and 280-303 aa in pUL40 could interact with pUL39, respectively. Deletion of 90-210 aa in pUL39 completely abrogated the interaction with pUL40. Deletion of 66-152, 218-258 and 280-303 aa in pUL40 remarkably weakened the interaction with pUL39, whereas a weak interaction could still be observed. Amino acid sequence alignments showed that the interaction domains identified in PRV pUL39/pUL40 were relatively non-conserved among the selected RNR subunits in alphaherpesviruses HSV1, HSV2, HHV3(VZV), BHV1, EHV1 and DEV. However, they were relatively conserved among PRV, HSV1 and HSV2. Collectively, our findings provided some molecular targets for inhibition of pUL39-pUL40 interaction to antagonize viral replication in PRV infected hosts.
Identifiants
pubmed: 32605757
pii: S0378-1135(20)30222-4
doi: 10.1016/j.vetmic.2020.108740
pii:
doi:
Substances chimiques
Protein Subunits
0
Ribonucleotide Reductases
EC 1.17.4.-
Nucleotidases
EC 3.1.3.-
deoxynucleoside-5'-monophosphatase
EC 3.1.3.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
108740Informations de copyright
Copyright © 2020 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare no competing interests.