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Structure paper

タイトル	Structures of influenza A virus RNA polymerase offer insight into viral genome replication.
ジャーナル・号・ページ	Nature, Vol. 573, Issue 7773, Page 287-290, Year 2019
掲載日	2019年9月4日
著者	Haitian Fan / Alexander P Walker / Loïc Carrique / Jeremy R Keown / Itziar Serna Martin / Dimple Karia / Jane Sharps / Narin Hengrung / Els Pardon / Jan Steyaert / Jonathan M Grimes / Ervin Fodor /
PubMed 要旨	Influenza A viruses are responsible for seasonal epidemics, and pandemics can arise from the transmission of novel zoonotic influenza A viruses to humans. Influenza A viruses contain a segmented ...Influenza A viruses are responsible for seasonal epidemics, and pandemics can arise from the transmission of novel zoonotic influenza A viruses to humans. Influenza A viruses contain a segmented negative-sense RNA genome, which is transcribed and replicated by the viral-RNA-dependent RNA polymerase (FluPol) composed of PB1, PB2 and PA subunits. Although the high-resolution crystal structure of FluPol of bat influenza A virus has previously been reported, there are no complete structures available for human and avian FluPol. Furthermore, the molecular mechanisms of genomic viral RNA (vRNA) replication-which proceeds through a complementary RNA (cRNA) replicative intermediate, and requires oligomerization of the polymerase-remain largely unknown. Here, using crystallography and cryo-electron microscopy, we determine the structures of FluPol from human influenza A/NT/60/1968 (H3N2) and avian influenza A/duck/Fujian/01/2002 (H5N1) viruses at a resolution of 3.0-4.3 Å, in the presence or absence of a cRNA or vRNA template. In solution, FluPol forms dimers of heterotrimers through the C-terminal domain of the PA subunit, the thumb subdomain of PB1 and the N1 subdomain of PB2. The cryo-electron microscopy structure of monomeric FluPol bound to the cRNA template reveals a binding site for the 3' cRNA at the dimer interface. We use a combination of cell-based and in vitro assays to show that the interface of the FluPol dimer is required for vRNA synthesis during replication of the viral genome. We also show that a nanobody (a single-domain antibody) that interferes with FluPol dimerization inhibits the synthesis of vRNA and, consequently, inhibits virus replication in infected cells. Our study provides high-resolution structures of medically relevant FluPol, as well as insights into the replication mechanisms of the viral RNA genome. In addition, our work identifies sites in FluPol that could be targeted in the development of antiviral drugs.
リンク	Nature / PubMed:31485076 / PubMed Central
手法	EM (単粒子) / X線回折
解像度	3.01 - 4.34 Å
構造データ	4660 6qwl EMDB-4660, PDB-6qwl: Influenza B virus (B/Panama/45) polymerase Hetermotrimer in complex with 3'5' cRNA promoter 手法: EM (単粒子) / 解像度: 4.1 Å 4661 6qx3 EMDB-4661, PDB-6qx3: Influenza A virus (A/NT/60/1968) polymerase Hetermotrimer in complex with 3'5' cRNA promoter and Nb8205 手法: EM (単粒子) / 解像度: 3.79 Å 4663 6qx8 EMDB-4663, PDB-6qx8: Influenza A virus (A/NT/60/1968) polymerase dimer of heterotrimer in complex with 5' cRNA promoter 手法: EM (単粒子) / 解像度: 4.07 Å EMDB-4664: Influenza A virus (A/NT/60/1968) polymerase dimer of Hetermotrimer in complex with 3'5' cRNA promoter 手法: EM (単粒子) / 解像度: 4.34 Å 4666 6qxe EMDB-4666, PDB-6qxe: Influenza A virus (A/NT/60/1968) polymerase dimer of hetermotrimer in complex with 3'5' cRNA promoter and Nb8205 手法: EM (単粒子) / 解像度: 4.15 Å 4986 6rr7 EMDB-4986, PDB-6rr7: Influenza A virus (A/NT/60/1968) polymerase Heterotrimer bound to 3'5' vRNA promoter and capped RNA primer 手法: EM (単粒子) / 解像度: 3.01 Å PDB-6qnw: Influenza A Polymerase Heterotrimer Human H3N2 Northern Territory 1968 手法: X-RAY DIFFRACTION / 解像度: 3.31 Å PDB-6qpf: Influenza A virus Polymerase Heterotrimer A/duck/Fujian/01/2002(H5N1) 手法: X-RAY DIFFRACTION / 解像度: 3.634 Å PDB-6qpg: Influenza A virus Polymerase Heterotrimer A/nt/60/1968(H3N2) in complex with Nanobody NB8205 手法: X-RAY DIFFRACTION / 解像度: 3.34 Å
由来	Influenza B virus (B/Panama/45/1990) (B型インフルエンザウイルス) influenza a virus (A型インフルエンザウイルス) influenza b virus (strain b/panama/45/1990) (B型インフルエンザウイルス) influenza a virus (a/nt/60/1968(h3n2)) (A型インフルエンザウイルス) lama glama (ラマ) influenza a virus (strain a/hong kong/1/1968 h3n2) (A型インフルエンザウイルス) influenza a virus (strain a/northern territory/60/1968 h3n2) (A型インフルエンザウイルス) Influenza A virus (A/nt/60/1968(H3N2)) spodoptera frugiperda (ツマジロクサヨトウ) influenza a virus (a/northern territory/60/1968(h3n2)) (A型インフルエンザウイルス) camelidae (ラクダ科)
キーワード	RNA BINDING PROTEIN (RNA結合タンパク質) / Influenza A (A型インフルエンザウイルス) / RNA polymerase (RNAポリメラーゼ) / Influenza polymerase / Influenza dimer / RDRP (RNA依存性RNAポリメラーゼ) / Nanobody (ナノボディ) / antiviral inhibitor