Characterization of Two Evolutionarily Conserved, Alternatively Spliced Nuclear Phosphoproteins, NFAR-1 and -2, that Function in mRNA Processing and Interact with the Double-stranded RNA-dependent Protein Kinase, PKR

Laura R. Saunders, Darren J. Perkins, Siddharth Balachandran, Rebecca Michaels, Ryan Ford, Akila Mayeda, Glen N Barber

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

We report here the isolation and characterization of two proteins, NFAR-1 and -2, which were isolated through their ability to interact with the dsRNA-dependent protein kinase, PKR. The NFAR proteins, of 90 and 110 kDa, are derived from a single gene through alternative splicing and are evolutionarily conserved nuclear phosphoproteins that interact with double-stranded RNA. Both NFAR-1 and -2 are phosphorylated by PKR, reciprocally co-immunoprecipitate with PKR, and colocalize with the kinase in a diffuse nuclear pattern within the cell. Transfection studies indicate that the NFARs regulate gene expression at the level of transcription, probably during the processing of pre-mRNAs, an activity that was increased in fibroblasts lacking PKR. Subsequent functional analyses indicated that amino acids important for NFAR's activity were localized to the C terminus of the protein, a region that was found to specifically interact with FUS and SMN, proteins also known as regulators of RNA processing. Accordingly, both NFARs were found to associate with both pre-mRNAs and spliced mRNAs in post-transcriptional studies, similar to the known splicing factor ASF/ SF-2. Collectively, our data indicate that the NFARs may facilitate double-stranded RNA-regulated gene expression at the level of post-transcription and possibly contribute to host defense-related mechanisms in the cell.

Original languageEnglish
Pages (from-to)32300-32312
Number of pages13
JournalJournal of Biological Chemistry
Volume276
Issue number34
DOIs
StatePublished - Aug 24 2001

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eIF-2 Kinase
Double-Stranded RNA
Phosphoproteins
RNA Precursors
RNA-Binding Protein FUS
Gene Expression
Messenger RNA
Alternative Splicing
Transcription
Processing
Protein C
Gene expression
Transfection
Proteins
Phosphotransferases
Fibroblasts
RNA
Amino Acids
Genes
RNA Splicing Factors

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of Two Evolutionarily Conserved, Alternatively Spliced Nuclear Phosphoproteins, NFAR-1 and -2, that Function in mRNA Processing and Interact with the Double-stranded RNA-dependent Protein Kinase, PKR. / Saunders, Laura R.; Perkins, Darren J.; Balachandran, Siddharth; Michaels, Rebecca; Ford, Ryan; Mayeda, Akila; Barber, Glen N.

In: Journal of Biological Chemistry, Vol. 276, No. 34, 24.08.2001, p. 32300-32312.

Research output: Contribution to journalArticle

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