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Journal of Virology, January 2000, p. 669-675, Vol. 74, No. 2
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Profilin Is Required for Optimal Actin-Dependent Transcription of Respiratory Syncytial Virus Genome RNA

Emily Burke,1 Nicole M. Mahoney,2 Steven C. Almo,2 and Sailen Barik1,*

Department of Biochemistry and Molecular Biology, University of South Alabama College of Medicine, Mobile, Alabama 36688-0002,1 and Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York 104612

Received 13 July 1999/Accepted 14 October 1999

Transcription of human respiratory syncytial virus (RSV) genome RNA exhibited an obligatory need for the host cytoskeletal protein actin. Optimal transcription, however, required the participation of another cellular protein that was characterized as profilin by a number of criteria. The amino acid sequence of the protein, purified on the basis of its transcription-optimizing activity in vitro, exactly matched that of profilin. RSV transcription was inhibited 60 to 80% by antiprofilin antibody or poly-L-proline, molecules that specifically bind profilin. Native profilin, purified from extracts of lung epithelial cells by affinity binding to a poly-L-proline matrix, stimulated the actin-saturated RSV transcription by 2.5- to 3-fold. Recombinant profilin, expressed in bacteria, stimulated viral transcription as effectively as the native protein and was also inhibited by poly-L-proline. Profilin alone, in the absence of actin, did not activate viral transcription. It is estimated that at optimal levels of transcription, every molecule of viral genomic RNA associates with approximately the following number of protein molecules: 30 molecules of L, 120 molecules of phosphoprotein P, and 60 molecules each of actin and profilin. Together, these results demonstrated for the first time a cardinal role for profilin, an actin-modulatory protein, in the transcription of a paramyxovirus RNA genome.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, University of South Alabama College of Medicine, 307 University Blvd., Mobile, AL 36688-0002. Phone: (334) 460-6860. Fax: (334) 460-6865. E-mail: sbarik{at}jaguar1.usouthal.edu.

Journal of Virology, January 2000, p. 669-675, Vol. 74, No. 2
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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