Flexibility of NS5 Methyltransferase-Polymerase Linker Region Is Essential for Dengue Virus Replication.
We examined the function of the conserved Val/Ile residue within the dengue virus NS5 interdomain linker (residues 263 to 272) by site-directed mutagenesis. Gly substitution or Gly/Pro insertion after the conserved residue increased the linker flexibility and created slightly attenuated viruses. In contrast, Pro substitution abolished virus replication by imposing rigidity in the linker and restricting NS5's conformational plasticity. Our biochemical and reverse genetics experiments demonstrate that NS5 utilizes conformational regulation to achieve optimum viral replication.
Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore.
Yongqian
Y
Zhao
School of Biological Sciences, Nanyang Technological University, Singapore Novartis Institute for Tropical Diseases, Singapore.
Tingjin Sherryl
TS
Soh
Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
Kitti Wing Ki
KW
Chan
Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore.
Sarah Suet Yin
SS
Fung
NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore Department of Biological Sciences, National University of Singapore, Singapore.
Kunchithapadam
K
Swaminathan
Novartis Institute for Tropical Diseases, Singapore.
Siew Pheng
SP
Lim
Novartis Institute for Tropical Diseases, Singapore.
Pei-Yong
PY
Shi
Research School of Chemistry, Australian National University, Canberra, Australia.
Thomas
T
Huber
School of Biological Sciences, Nanyang Technological University, Singapore UPMC UMRS CR7-CNRS ERL 8255-INSERM U1135 Centre d'Immunologie et des Maladies Infectieuses, Centre Hospitalier Universitaire Pitié-Salpêtrière, Faculté de Médecine Pierre et Marie Curie, Paris, France.
Julien
J
Lescar
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore luodahai@ntu.edu.sg subhash.vasudevan@duke-nus.edu.sg.
Dahai
D
Luo
Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, Singapore NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore luodahai@ntu.edu.sg subhash.vasudevan@duke-nus.edu.sg.
Subhash G
SG
Vasudevan
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003412:Cricetulus; D018360:Crystallography, X-Ray; D003716:Dengue Virus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009154:Mutation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000502:physiology*
C582223:NS5 protein, dengue virus; D012367:RNA, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins