RRC ID 69999
Author Hu Y, Gor V, Morikawa K, Nagata K, Kawaguchi A.
Title Cellular splicing factor UAP56 stimulates trimeric NP formation for assembly of functional influenza viral ribonucleoprotein complexes.
Journal Sci Rep
Abstract The influenza virus RNA genome exists as a ribonucleoprotein (RNP) complex by interacting with NP, one of virus-encoded RNA binding proteins. It is proposed that trimeric NP is a functional form, but it is not clear how trimeric NP is formed and transferred to RNA. UAP56, a cellular splicing factor, functions as a molecular chaperone for NP and is required for the replication-coupled RNP formation of newly synthesized viral genome, but the details of NP transfer to viral RNA by UAP56 is unclear. Here we found that UAP56 is complexed with trimeric NP, but not monomeric NP. Gel filtration analysis and atomic force microscopy analysis indicated that the complex consists of two trimeric NP connected by UAP56. We also found that UAP56 stimulates trimeric NP formation from monomeric NP even at physiological salt concentrations. Thus, UAP56 facilitates the transfer of NP to viral RNA since trimeric NP has higher RNA binding activity than monomeric NP. Further, UAP56 represses the binding of excess amount of NP to RNA possibly by transferring trimeric NP. Collectively, we propose that UAP56 stimulates viral RNP formation through promotion of the assembly of trimeric NP and is important for the structural integrity of NP-RNA complex.
Volume 7(1)
Pages 14053
Published 2017-10-25
DOI 10.1038/s41598-017-13784-4
PII 10.1038/s41598-017-13784-4
PMID 29070793
PMC PMC5656576
MeSH DEAD-box RNA Helicases / genetics DEAD-box RNA Helicases / metabolism* Gene Expression Regulation, Viral* Genome, Viral Humans Influenza A virus / genetics Influenza A virus / metabolism* Influenza, Human / genetics Influenza, Human / metabolism* Influenza, Human / virology Protein Binding RNA Splicing RNA, Viral / genetics RNA, Viral / metabolism* RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism* Ribonucleoproteins / genetics Ribonucleoproteins / metabolism*
IF 3.998
DNA material His-tagged NP on pET14b (RDB19458)