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
N1 - Funding Information:
This work was supported by National Research Foundation Grants funded by the Korean Government (2012R1A1A2004532 and 2018R1A2B6001363 to M. J. Song and 2017R1A2B2005666 and 2018R1A44A1022589 to K. Y. Hwang).
Publisher Copyright:
© 2018 Woo-Chang Chung et al.
PY - 2018
Y1 - 2018
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 - X-ray crystallography
KW - murine gammaherpesvirus 68
KW - open reading frame 49
KW - poly(ADP-ribose) polymerase 1
KW - structure determination
KW - viral PARP-1-interacting protein
UR - http://www.scopus.com/inward/record.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 -