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Journal of Virology, September 1999, p. 7255-7261, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Moloney Murine Sarcoma Virus Genomic RNAs Dimerize via a Two-Step Process: a Concentration-Dependent Kissing-Loop Interaction Is Driven by Initial Contact between Consecutive Guanines

Hinh Ly,1 Donald P. Nierlich,2,3 John C. Olsen,4,5 and Andrew H. Kaplan1,4,6,*

Departments of Microbiology & Immunology1 and Medicine,4 Lineberger Cancer Center,6 and Cystic Fibrosis/Pulmonary Research and Treatment Center,5 School of Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, and Department of Microbiology, Immunology, & Molecular Genetics2 and Molecular Biology Institute,3 University of California Los Angeles School of Medicine, Los Angeles, California

Received 10 February 1999/Accepted 25 May 1999

Retroviruses contain two plus-strand genomic RNAs, which are stably but noncovalently joined in their 5' regions by a dimer linkage structure (DLS). Two models have been put forward to explain the mechanisms by which the RNAs dimerize; each model emphasizes the role of specific molecular determinants. The kissing-loop model implicates interactions between palindromic sequences in the DLS region. The second model proposes that purine-rich stretches in the region form purine quartet structures. Here, we present an examination of the in vitro dimerization of Moloney murine sarcoma virus (MuSV) RNA in the context of these two models. Dimers were found to form spontaneously in a temperature-, time-, concentration-, and salt-dependent manner. In contrast to earlier reports, we found that deletion of neither the palindrome nor the consensus purine motifs (PuGGAPuA) affected the level of dimer formation at low concentrations of RNA. Rather, different purine-rich sequences, i.e., consecutive stretches of guanines, were found to enhance both in vitro RNA dimerization and in vivo viral replication. Biochemical evidence further suggests that these guanine-rich (G-rich) stretches form guanine quartet structures. We also found that the palindromic sequences could support dimerization at significantly higher RNA concentrations. In addition, the G-rich stretches were as important as the palindromic sequence for maintaining efficient viral replication. Overall, our data support a model that entails contributions from both of the previously proposed mechanisms of retroviral RNA dimerization.

* Corresponding author. Mailing address: CB#7030, 547 Burnett-Womack, UNC-Chapel Hill, Chapel Hill, NC 27599-7030. Phone: (919) 966-2536. Fax: (919) 966-6714. E-mail: akaplan{at}med.unc.edu.

Journal of Virology, September 1999, p. 7255-7261, Vol. 73, No. 9
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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