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Journal of Virology, January 1999, p. 772-777, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Polypyrimidine Tract-Binding Protein Binds to the Leader RNA of Mouse Hepatitis Virus and Serves as a Regulator of Viral Transcription

Hsin-Pai Li,1 Peiyong Huang,1 Sungmin Park,1 and Michael M. C. Lai1,2,*

Department of Molecular Microbiology and Immunology1 and Howard Hughes Medical Institute,2 University of Southern California School of Medicine, Los Angeles, California 90033-1054

Received 29 April 1998/Accepted 25 September 1998

A cellular protein, previously described as p55, binds specifically to the plus strand of the mouse hepatitis virus (MHV) leader RNA. We have purified this protein and determined by partial peptide sequencing that it is polypyrimidine tract-binding protein (PTB) (also known as heterogeneous nuclear ribonucleoprotein [hnRNP] I), a nuclear protein which shuttles between the nucleus and cytoplasm. PTB plays a role in the regulation of alternative splicing of pre-mRNAs in normal cells and translation of several viruses. By UV cross-linking and immunoprecipitation studies using cellular extracts and a recombinant PTB, we have established that PTB binds to the MHV plus-strand leader RNA specifically. Deletion analyses of the leader RNA mapped the PTB-binding site to the UCUAA pentanucleotide repeats. Using a defective-interfering RNA reporter system, we have further shown that the PTB-binding site in the leader RNA is critical for MHV RNA synthesis. This and our previous study (H.-P. Li, X. Zhang, R. Duncan, L. Comai, and M. M. C. Lai, Proc. Natl. Acad. Sci. USA 94:9544-9549, 1997) combined thus show that two cellular hnRNPs, PTB and hnRNP A1, bind to the transcription-regulatory sequences of MHV RNA and may participate in its transcription.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, Department of Molecular Microbiology and Immunology, University of Southern California School of Medicine, 2011 Zonal Ave., HMR-401, Los Angeles, CA 90033-1054. Phone: (323) 442-1748. Fax: (323) 442-9555. E-mail: michlai{at}hsc.usc.edu.

Journal of Virology, January 1999, p. 772-777, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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