This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hibbert, C. S. Articles by Rein, A. Search for Related Content PubMed PubMed Citation Articles by Hibbert, C. S. Articles by Rein, A.

 Previous Article  |  Next Article 

Journal of Virology, July 2005, p. 8142-8148, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8142-8148.2005

Preliminary Physical Mapping of RNA-RNA Linkages in the Genomic RNA of Moloney Murine Leukemia Virus

HIV Drug Resistance Program, National Cancer Institute—Frederick, Frederick, Maryland 21702-1201

Received 7 December 2004/ Accepted 16 March 2005

Retrovirus particles contain two copies of their genomic RNA, held together in a dimer by linkages which presumably consist of a limited number of base pairs. In an effort to localize these linkages, we digested deproteinized RNA from Moloney murine leukemia virus (MLV) particles with RNase H in the presence of oligodeoxynucleotides complementary to specific sites in viral RNA. The cleaved RNAs were then characterized by nondenaturing gel electrophoresis. We found that fragments composed of nucleotides 1 to 754 were dimeric, with a linkage as thermostable as that between dimers of intact genomic RNA. In contrast, there was no stable linkage between fragments consisting of nucleotides 755 to 8332. Thus, the most stable linkage between monomers is on the 5' side of nucleotide 754. This conclusion is in agreement with earlier electron microscopic analyses of partially denatured viral RNAs and with our study (C. S. Hibbert, J. Mirro, and A. Rein, J. Virol. 78:10927-10938, 2004) of encapsidated nonviral mRNAs containing inserts of viral sequence. We obtained similar results with RNAs from immature MLV particles, in which the dimeric linkage is different from that in mature particles and has not previously been localized. The 5' and 3' fragments of cleaved RNA are all held together by thermolabile linkages, indicating the presence of tethering interactions between bases 5' and bases 3' of the cleavage site. When RNAs from mature particles were cleaved at nucleotide 1201, we detected tethering interactions spanning the cleavage site which are intramonomeric and are as strong as the most stable linkage between the monomers.

This article has been cited by other articles:

• Rulli, S. J. Jr., Hibbert, C. S., Mirro, J., Pederson, T., Biswal, S., Rein, A. (2007). Selective and Nonselective Packaging of Cellular RNAs in Retrovirus Particles. J. Virol. 81: 6623-6631 [Abstract] [Full Text]  
• Gherghe, C., Weeks, K. M. (2006). The SL1-SL2 (Stem-Loop) Domain Is the Primary Determinant for Stability of the Gamma Retroviral Genomic RNA Dimer. J. Biol. Chem. 281: 37952-37961 [Abstract] [Full Text]  
• Kasprzak, W., Bindewald, E., Shapiro, B. A. (2005). Structural polymorphism of the HIV-1 leader region explored by computational methods. Nucleic Acids Res 33: 7151-7163 [Abstract] [Full Text]