TY - JOUR
T1 - Critical phosphoprotein elements that regulate polymerase architecture and function in vesicular stomatitis virus
AU - Rahmeh, Amal A.
AU - Morin, Benjamin
AU - Schenk, Andreas D.
AU - Liang, Bo
AU - Heinrich, Bianca S.
AU - Brusic, Vesna
AU - Walz, Thomas
AU - Whelan, Sean P.J.
PY - 2012/9/4
Y1 - 2012/9/4
N2 - The RNA-dependent RNA polymerase (RdRP) of nonsegmented negative-sense RNA viruses consists of a large catalytic protein (L) and a phosphoprotein cofactor (P). During infection, the RdRP replicates and transcribes the viral genome, which resides inside an oligomer of nucleocapsid protein (N-RNA). The classical view of P as a cofactor for L assigns a primary role of P as a bridge mediating the access of L to the RNA template, whereby its N-terminal domain (PNTD) binds L and its C-terminal domain (PCTD) binds NRNA. Recent biochemical and structural studies of a prototype nonsegmented negative-sense RNA virus, vesicular stomatitis virus, suggest a role for P beyond that of a mere physical link: P induces a structural rearrangement in L and stimulates polymerase processivity. In this study, we investigated the critical requirements within P mediating the functional interaction with L to form a fully functional RdRP. We analyzed the correlation between the impact of P on the conformation of L and its activity in RNA synthesis and the consequences of these events on RdRP function. We identified three separable elements of the PNTD that are required for inducing the conformational rearrangement of L, stimulating polymerase processivity, and mediating transcription of the NRNA. The functional interplay between these elements provides insight into the role of P as a dynamic player in the RNA synthesis machine, influencing essential aspects of polymerase structure and function.
AB - The RNA-dependent RNA polymerase (RdRP) of nonsegmented negative-sense RNA viruses consists of a large catalytic protein (L) and a phosphoprotein cofactor (P). During infection, the RdRP replicates and transcribes the viral genome, which resides inside an oligomer of nucleocapsid protein (N-RNA). The classical view of P as a cofactor for L assigns a primary role of P as a bridge mediating the access of L to the RNA template, whereby its N-terminal domain (PNTD) binds L and its C-terminal domain (PCTD) binds NRNA. Recent biochemical and structural studies of a prototype nonsegmented negative-sense RNA virus, vesicular stomatitis virus, suggest a role for P beyond that of a mere physical link: P induces a structural rearrangement in L and stimulates polymerase processivity. In this study, we investigated the critical requirements within P mediating the functional interaction with L to form a fully functional RdRP. We analyzed the correlation between the impact of P on the conformation of L and its activity in RNA synthesis and the consequences of these events on RdRP function. We identified three separable elements of the PNTD that are required for inducing the conformational rearrangement of L, stimulating polymerase processivity, and mediating transcription of the NRNA. The functional interplay between these elements provides insight into the role of P as a dynamic player in the RNA synthesis machine, influencing essential aspects of polymerase structure and function.
KW - Large polymerase
KW - Mononegavirales
KW - Replication and transcription
KW - Rhabdovirus
UR - http://www.scopus.com/inward/record.url?scp=84865963708&partnerID=8YFLogxK
U2 - 10.1073/pnas.1209147109
DO - 10.1073/pnas.1209147109
M3 - Article
C2 - 22908284
AN - SCOPUS:84865963708
SN - 0027-8424
VL - 109
SP - 14628
EP - 14633
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 36
ER -