TY - JOUR
T1 - siRNA screen of early poxvirus genes identifies the AAA+ ATPase D5 as the virus genome-uncoating factor
AU - Kilcher, Samuel
AU - Schmidt, Florian Ingo
AU - Schneider, Christoph
AU - Kopf, Manfred
AU - Helenius, Ari
AU - Mercer, Jason
PY - 2014/1/15
Y1 - 2014/1/15
N2 - Summary Poxvirus genome uncoating is a two-step process. First, cytoplasmic viral cores are activated and early viral genes are expressed. Next, cores are disassembled and the genomes released. This second step depends on an early viral factor(s) that has eluded identification for over 40 years. We used a large-scale, high-throughput RNAi screen directed against vaccinia virus (VACV) to identify the VACV AAA+ ATPase D5 as the poxvirus uncoating factor. We show that the ATPase activity of D5 is required for uncoating. Superresolution microscopy suggests that D5 acts directly at viral cores for genome release. Thus, the putative helicase D5 is a multifunctional protein required for genome uncoating and replication. Additionally, in vivo delivery of anti-D5 siRNAs reduced virus production in a mouse model of VACV infection. These results demonstrate the use of virus-targeting RNAi libraries to investigate viral gene function and suggest therapeutic avenues.
AB - Summary Poxvirus genome uncoating is a two-step process. First, cytoplasmic viral cores are activated and early viral genes are expressed. Next, cores are disassembled and the genomes released. This second step depends on an early viral factor(s) that has eluded identification for over 40 years. We used a large-scale, high-throughput RNAi screen directed against vaccinia virus (VACV) to identify the VACV AAA+ ATPase D5 as the poxvirus uncoating factor. We show that the ATPase activity of D5 is required for uncoating. Superresolution microscopy suggests that D5 acts directly at viral cores for genome release. Thus, the putative helicase D5 is a multifunctional protein required for genome uncoating and replication. Additionally, in vivo delivery of anti-D5 siRNAs reduced virus production in a mouse model of VACV infection. These results demonstrate the use of virus-targeting RNAi libraries to investigate viral gene function and suggest therapeutic avenues.
UR - http://www.scopus.com/inward/record.url?scp=84892621131&partnerID=8YFLogxK
U2 - 10.1016/j.chom.2013.12.008
DO - 10.1016/j.chom.2013.12.008
M3 - Article
C2 - 24439902
AN - SCOPUS:84892621131
SN - 1931-3128
VL - 15
SP - 103
EP - 112
JO - Cell Host and Microbe
JF - Cell Host and Microbe
IS - 1
ER -