Abstract
Poly(ADP-ribose) polymerase 1 (PARP-1), an enzyme that modifies nuclear proteins by poly(ADP-ribosyl)ation, regulates various cellular activities and restricts the lytic replication of oncogenic gammaherpesviruses by inhibiting the function of replication and transcription activator (RTA), a key switch molecule of the viral life cycle. A viral PARP-1-interacting protein (vPIP) encoded by murine gammaherpesvirus 68 (MHV-68) orf49 facilitates lytic replication by disrupting interactions between PARP-1 and RTA. Here, the structure of MHV-68 vPIP was determined at 2.2 Å resolution. The structure consists of 12 α-helices with characteristic N-terminal β-strands (Nβ) and forms a V-shaped-twist dimer in the asymmetric unit. Structure-based mutagenesis revealed that Nβ and the α1 helix (residues 2-26) are essential for the nuclear localization and function of vPIP; three residues were then identified (Phe5, Ser12 and Thr16) that were critical for the function of vPIP and its interaction with PARP-1. A recombinant MHV-68 harboring mutations of these three residues showed severely attenuated viral replication both in vitro and in vivo. Moreover, ORF49 of Kaposi's sarcoma-associated herpesvirus also directly interacted with PARP-1, indicating a conserved mechanism of action of vPIPs. The results elucidate the novel molecular mechanisms by which oncogenic gammaherpesviruses overcome repression by PARP-1 using vPIPs.
| Original language | English |
|---|---|
| Pages (from-to) | 866-879 |
| Number of pages | 14 |
| Journal | IUCrJ |
| Volume | 5 |
| DOIs | |
| Publication status | Published - 2018 Jan 1 |
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Keywords
- Kaposi's sarcoma-associated herpesvirus
- murine gammaherpesvirus 68
- open reading frame 49
- poly(ADP-ribose) polymerase 1
- structure determination
- viral PARP-1-interacting protein
- X-ray crystallography
ASJC Scopus subject areas
- Chemistry(all)
- Biochemistry
- Materials Science(all)
- Condensed Matter Physics
Cite this
Structure-based mechanism of action of a viral poly(ADP-ribose) polymerase 1-interacting protein facilitating virus replication. / Chung, Woo Chang; Kim, Junsoo; Kim, Byung Chul; Kang, Hye Ri; Son, Jonghyeon; Ki, Hosam; Hwang, Kwang Yeon; Song, Moon Jung.
In: IUCrJ, Vol. 5, 01.01.2018, p. 866-879.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Structure-based mechanism of action of a viral poly(ADP-ribose) polymerase 1-interacting protein facilitating virus replication
AU - Chung, Woo Chang
AU - Kim, Junsoo
AU - Kim, Byung Chul
AU - Kang, Hye Ri
AU - Son, Jonghyeon
AU - Ki, Hosam
AU - Hwang, Kwang Yeon
AU - Song, Moon Jung
PY - 2018/1/1
Y1 - 2018/1/1
N2 - Poly(ADP-ribose) polymerase 1 (PARP-1), an enzyme that modifies nuclear proteins by poly(ADP-ribosyl)ation, regulates various cellular activities and restricts the lytic replication of oncogenic gammaherpesviruses by inhibiting the function of replication and transcription activator (RTA), a key switch molecule of the viral life cycle. A viral PARP-1-interacting protein (vPIP) encoded by murine gammaherpesvirus 68 (MHV-68) orf49 facilitates lytic replication by disrupting interactions between PARP-1 and RTA. Here, the structure of MHV-68 vPIP was determined at 2.2 Å resolution. The structure consists of 12 α-helices with characteristic N-terminal β-strands (Nβ) and forms a V-shaped-twist dimer in the asymmetric unit. Structure-based mutagenesis revealed that Nβ and the α1 helix (residues 2-26) are essential for the nuclear localization and function of vPIP; three residues were then identified (Phe5, Ser12 and Thr16) that were critical for the function of vPIP and its interaction with PARP-1. A recombinant MHV-68 harboring mutations of these three residues showed severely attenuated viral replication both in vitro and in vivo. Moreover, ORF49 of Kaposi's sarcoma-associated herpesvirus also directly interacted with PARP-1, indicating a conserved mechanism of action of vPIPs. The results elucidate the novel molecular mechanisms by which oncogenic gammaherpesviruses overcome repression by PARP-1 using vPIPs.
AB - Poly(ADP-ribose) polymerase 1 (PARP-1), an enzyme that modifies nuclear proteins by poly(ADP-ribosyl)ation, regulates various cellular activities and restricts the lytic replication of oncogenic gammaherpesviruses by inhibiting the function of replication and transcription activator (RTA), a key switch molecule of the viral life cycle. A viral PARP-1-interacting protein (vPIP) encoded by murine gammaherpesvirus 68 (MHV-68) orf49 facilitates lytic replication by disrupting interactions between PARP-1 and RTA. Here, the structure of MHV-68 vPIP was determined at 2.2 Å resolution. The structure consists of 12 α-helices with characteristic N-terminal β-strands (Nβ) and forms a V-shaped-twist dimer in the asymmetric unit. Structure-based mutagenesis revealed that Nβ and the α1 helix (residues 2-26) are essential for the nuclear localization and function of vPIP; three residues were then identified (Phe5, Ser12 and Thr16) that were critical for the function of vPIP and its interaction with PARP-1. A recombinant MHV-68 harboring mutations of these three residues showed severely attenuated viral replication both in vitro and in vivo. Moreover, ORF49 of Kaposi's sarcoma-associated herpesvirus also directly interacted with PARP-1, indicating a conserved mechanism of action of vPIPs. The results elucidate the novel molecular mechanisms by which oncogenic gammaherpesviruses overcome repression by PARP-1 using vPIPs.
KW - Kaposi's sarcoma-associated herpesvirus
KW - murine gammaherpesvirus 68
KW - open reading frame 49
KW - poly(ADP-ribose) polymerase 1
KW - structure determination
KW - viral PARP-1-interacting protein
KW - X-ray crystallography
UR - http://www.scopus.com/inward/record.url?scp=85056186175&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85056186175&partnerID=8YFLogxK
U2 - 10.1107/S2052252518013854
DO - 10.1107/S2052252518013854
M3 - Article
AN - SCOPUS:85056186175
VL - 5
SP - 866
EP - 879
JO - IUCrJ
JF - IUCrJ
SN - 2052-2525
ER -