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Journal of Virology, June 2007, p. 6623-6631, Vol. 81, No. 12
0022-538X/07/$08.00+0     doi:10.1128/JVI.02833-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Selective and Nonselective Packaging of Cellular RNAs in Retrovirus Particles{triangledown}

Samuel J. Rulli Jr.,1 Catherine S. Hibbert,1 Jane Mirro,1 Thoru Pederson,2 Shyam Biswal,3 and Alan Rein1*

HIV Drug Resistance Program, National Cancer Institute, Frederick, Maryland 21702,1 Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605,2 Department of Environmental Health Sciences, Bloomberg School of Public Health, and Department of Oncology, School of Medicine, Johns Hopkins University, Baltimore, Maryland 212053

Received 21 December 2006/ Accepted 20 March 2007

Assembly of retrovirus particles normally entails the selective encapsidation of viral genomic RNA. However, in the absence of packageable viral RNA, assembly is still efficient, and the released virus-like particles (termed "{Psi}–" particles) still contain roughly normal amounts of RNA. We have proposed that cellular mRNAs replace the genome in {Psi}– particles. We have now analyzed the mRNA content of {Psi}– and {Psi}+ murine leukemia virus (MLV) particles using both microarray analysis and real-time reverse transcription-PCR. The majority of mRNA species present in the virus-producing cells were also detected in {Psi}– particles. Remarkably, nearly all of them were packaged nonselectively; that is, their representation in the particles was simply proportional to their representation in the cells. However, a small number of low-abundance mRNAs were greatly enriched in the particles. In fact, one mRNA species was enriched to the same degree as {Psi}+ genomic RNA. Similar results were obtained with particles formed from the human immunodeficiency virus type 1 (HIV-1) Gag protein, and the same mRNAs were enriched in MLV and HIV-1 particles. The levels of individual cellular mRNAs were ~5- to 10-fold higher in {Psi}– than in {Psi}+ MLV particles, in agreement with the idea that they are replacing viral RNA in the former. In contrast, signal recognition particle RNA was present at the same level in {Psi}– and {Psi}+ particles; a minor fraction of this RNA was weakly associated with genomic RNA in {Psi}+ MLV particles.

* Corresponding author. Mailing address: National Cancer Institute—Frederick, P.O. Box B, Frederick, MD 21702-1201. Phone: (301) 846-1361. Fax: (301) 846-6013. E-mail: rein{at}ncifcrf.gov

{triangledown} Published ahead of print on 28 March 2007.

Journal of Virology, June 2007, p. 6623-6631, Vol. 81, No. 12
0022-538X/07/$08.00+0     doi:10.1128/JVI.02833-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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