Sztuba-Solinska Joanna, Diaz Larissa, Kumar Mia R, Kolb Gaëlle, Wiley Michael R, Jozwick Lucas, Kuhn Jens H, Palacios Gustavo, Radoshitzky Sheli R, J Le Grice Stuart F, Johnson Reed F
RT Biochemistry Section, Basic Research Laboratory, National Cancer Institute-Frederick, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA.
Emerging Viral Pathogens Section, National Institute of Allergy and Infectious Disease, National Institutes of Health, Fort Detrick, Frederick, MD 21702, USA.
Nucleic Acids Res. 2016 Nov 16;44(20):9831-9846. doi: 10.1093/nar/gkw825. Epub 2016 Sep 19.
Ebola virus (EBOV) is a single-stranded negative-sense RNA virus belonging to the Filoviridae family. The leader and trailer non-coding regions of the EBOV genome likely regulate its transcription, replication, and progeny genome packaging. We investigated the cis-acting RNA signals involved in RNA-RNA and RNA-protein interactions that regulate replication of eGFP-encoding EBOV minigenomic RNA and identified heat shock cognate protein family A (HSC70) member 8 (HSPA8) as an EBOV trailer-interacting host protein. Mutational analysis of the trailer HSPA8 binding motif revealed that this interaction is essential for EBOV minigenome replication. Selective 2'-hydroxyl acylation analyzed by primer extension analysis of the secondary structure of the EBOV minigenomic RNA indicates formation of a small stem-loop composed of the HSPA8 motif, a 3' stem-loop (nucleotides 1868-1890) that is similar to a previously identified structure in the replicative intermediate (RI) RNA and a panhandle domain involving a trailer-to-leader interaction. Results of minigenome assays and an EBOV reverse genetic system rescue support a role for both the panhandle domain and HSPA8 motif 1 in virus replication.
埃博拉病毒（EBOV）是一种属于丝状病毒科的单链负义RNA病毒。EBOV基因组的前导和尾随非编码区可能调控其转录、复制及子代基因组包装。我们研究了参与调控编码eGFP的EBOV微型基因组RNA复制的RNA-RNA和RNA-蛋白质相互作用中的顺式作用RNA信号，并鉴定出热休克同源蛋白家族A（HSC70）成员8（HSPA8）为一种与EBOV尾随序列相互作用的宿主蛋白。对尾随序列HSPA8结合基序的突变分析表明，这种相互作用对EBOV微型基因组复制至关重要。通过对EBOV微型基因组RNA二级结构进行引物延伸分析的选择性2'-羟基酰化表明，形成了一个由HSPA8基序、一个与复制中间体（RI）RNA中先前鉴定的结构相似的3'茎环（核苷酸1868 - 1890）以及一个涉及尾随到前导相互作用的锅柄结构域组成的小茎环。微型基因组分析结果和EBOV反向遗传系统拯救实验支持了锅柄结构域和HSPA8基序1在病毒复制中的作用。
Zotero 插件