A specific host cellular protein binding element near the 3' end of mouse hepatitis virus genomic RNA

Virology
Q LiuJ L Leibowitz

Abstract

A distinct host cellular protein binding element was mapped within a 38-nucleotide (nt) sequence 166-129 nucleotides upstream of the 3' end of the MHV-JHM genome using a RNase T1 protection/gel mobility shift electrophoresis assay. The resultant RNA-protein complex contains six host cellular proteins, one protein of 120-kDa molecular mass, two poorly resolved species approximately 55 kDa in size, a second pair of poorly resolved 40-kDa proteins, and a minor component of 25 kDa. A series of RNA probes containing deletions or clustered transversion mutations were tested for their ability to form complexes with mock- and MHV-JHM-infected cytoplasmic extracts. Three mutant RNA probes (mA, mB, and mC) with deletions at 154-140, 139-129, and 128-118, respectively, expressed 4, 37, and 94% of the host protein binding activity exhibited by the wild-type RNA. Defective interfering (DI) RNAs (DImA, DImB, and DImC) containing corresponding deletions at 154-140, 139-129, 128-118, and another DI RNA (DImD) with a deletion at nucleotides (nts) 112-102, a region which did not affect RNA-protein interactions, were transfected into MHV-JHM-infected 17CL-1 cells to assay the effects of these mutations on DI RNA replication. All of these mutation...Continue Reading

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Citations

Nov 12, 2005·Journal of Virology·Reed F JohnsonJulian L Leibowitz
Mar 5, 2015·Virus Research·Dong Yang, Julian L Leibowitz
Jan 27, 1999·Biochemical and Biophysical Research Communications·R T Chung, L M Kaplan

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