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A redox-sensitive cysteine in Zta is required for Epstein-Barr virus lytic cycle DNA replication.
J Virol. 2005 Nov; 79(21):13298-309.JV

Abstract

Epstein-Barr virus (EBV) reactivation from latency is known to be sensitive to redox regulation. The immediate-early protein Zta is a member of the basic-leucine zipper (bZIP) family of DNA binding proteins that stimulates viral and cellular transcription and nucleates a replication complex at the viral lytic origin. Zta shares with several members of the bZIP family a conserved cysteine residue (C189) that confers redox regulation of DNA binding. In this work, we show that replacement of C189 with serine (C189S) eliminated lytic cycle DNA replication function of Zta. The mechanistic basis for this replication defect was investigated. We show that C189S was not significantly altered for DNA binding activity in vitro or in vivo. We also show that C189S was not defective for transcription activation of EBV early gene promoters. C189S was deficient for transcription activation of several viral late genes that depend on lytic replication and therefore was consistent with a primary defect of C189S in activating lytic replication. C189S was not defective in binding methylated DNA binding sites and was capable of activating Rta from endogenous latent viral genomes, in contrast to the previously characterized S186A mutation. C189S was slightly impaired for its ability to form a stable complex with Rta, although this did not prevent Rta recruitment to OriLyt. C189S did provide some resistance to oxidation and nitrosylation, which potently inhibit Zta DNA binding activity in vitro. Interestingly, this redox sensitivity was not strictly dependent on C189S but involved additional cysteine residues in Zta. These results provide evidence that the conserved cysteine in the bZIP domain of Zta plays a primary role in EBV lytic cycle DNA replication.

Authors+Show Affiliations

The Wistar Institute, Philadelphia, PA 19104, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

16227252

Citation

Wang, Pu, et al. "A Redox-sensitive Cysteine in Zta Is Required for Epstein-Barr Virus Lytic Cycle DNA Replication." Journal of Virology, vol. 79, no. 21, 2005, pp. 13298-309.
Wang P, Day L, Dheekollu J, et al. A redox-sensitive cysteine in Zta is required for Epstein-Barr virus lytic cycle DNA replication. J Virol. 2005;79(21):13298-309.
Wang, P., Day, L., Dheekollu, J., & Lieberman, P. M. (2005). A redox-sensitive cysteine in Zta is required for Epstein-Barr virus lytic cycle DNA replication. Journal of Virology, 79(21), 13298-309.
Wang P, et al. A Redox-sensitive Cysteine in Zta Is Required for Epstein-Barr Virus Lytic Cycle DNA Replication. J Virol. 2005;79(21):13298-309. PubMed PMID: 16227252.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A redox-sensitive cysteine in Zta is required for Epstein-Barr virus lytic cycle DNA replication. AU - Wang,Pu, AU - Day,Latasha, AU - Dheekollu,Jayaraju, AU - Lieberman,Paul M, PY - 2005/10/18/pubmed PY - 2005/11/16/medline PY - 2005/10/18/entrez SP - 13298 EP - 309 JF - Journal of virology JO - J. Virol. VL - 79 IS - 21 N2 - Epstein-Barr virus (EBV) reactivation from latency is known to be sensitive to redox regulation. The immediate-early protein Zta is a member of the basic-leucine zipper (bZIP) family of DNA binding proteins that stimulates viral and cellular transcription and nucleates a replication complex at the viral lytic origin. Zta shares with several members of the bZIP family a conserved cysteine residue (C189) that confers redox regulation of DNA binding. In this work, we show that replacement of C189 with serine (C189S) eliminated lytic cycle DNA replication function of Zta. The mechanistic basis for this replication defect was investigated. We show that C189S was not significantly altered for DNA binding activity in vitro or in vivo. We also show that C189S was not defective for transcription activation of EBV early gene promoters. C189S was deficient for transcription activation of several viral late genes that depend on lytic replication and therefore was consistent with a primary defect of C189S in activating lytic replication. C189S was not defective in binding methylated DNA binding sites and was capable of activating Rta from endogenous latent viral genomes, in contrast to the previously characterized S186A mutation. C189S was slightly impaired for its ability to form a stable complex with Rta, although this did not prevent Rta recruitment to OriLyt. C189S did provide some resistance to oxidation and nitrosylation, which potently inhibit Zta DNA binding activity in vitro. Interestingly, this redox sensitivity was not strictly dependent on C189S but involved additional cysteine residues in Zta. These results provide evidence that the conserved cysteine in the bZIP domain of Zta plays a primary role in EBV lytic cycle DNA replication. SN - 0022-538X UR - https://www.unboundmedicine.com/medline/citation/16227252/A_redox_sensitive_cysteine_in_Zta_is_required_for_Epstein_Barr_virus_lytic_cycle_DNA_replication_ L2 - http://jvi.asm.org/cgi/pmidlookup?view=long&pmid=16227252 DB - PRIME DP - Unbound Medicine ER -